CN104813933B - Method for culturing yam seed tuber by using yam tissue culture seedling - Google Patents
Method for culturing yam seed tuber by using yam tissue culture seedling Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 33
- 238000012258 culturing Methods 0.000 title claims abstract description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 44
- 230000005540 biological transmission Effects 0.000 claims abstract description 6
- 244000061456 Solanum tuberosum Species 0.000 claims description 22
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 22
- 238000012360 testing method Methods 0.000 claims description 21
- 239000003337 fertilizer Substances 0.000 claims description 10
- 238000012549 training Methods 0.000 claims description 9
- 239000002689 soil Substances 0.000 claims description 7
- 239000000758 substrate Substances 0.000 claims description 7
- 239000003643 water by type Substances 0.000 claims description 6
- 241000196324 Embryophyta Species 0.000 claims description 5
- 230000008569 process Effects 0.000 claims description 5
- 239000005946 Cypermethrin Substances 0.000 claims description 3
- KAATUXNTWXVJKI-UHFFFAOYSA-N cypermethrin Chemical compound CC1(C)C(C=C(Cl)Cl)C1C(=O)OC(C#N)C1=CC=CC(OC=2C=CC=CC=2)=C1 KAATUXNTWXVJKI-UHFFFAOYSA-N 0.000 claims description 3
- 229960005424 cypermethrin Drugs 0.000 claims description 3
- 239000001963 growth medium Substances 0.000 claims description 3
- 230000003252 repetitive effect Effects 0.000 claims description 3
- 235000016709 nutrition Nutrition 0.000 claims description 2
- 230000035764 nutrition Effects 0.000 claims description 2
- 230000004083 survival effect Effects 0.000 abstract description 16
- 244000281702 Dioscorea villosa Species 0.000 abstract description 9
- 238000002054 transplantation Methods 0.000 abstract description 6
- 241000700605 Viruses Species 0.000 abstract description 2
- 238000012271 agricultural production Methods 0.000 abstract description 2
- 230000008859 change Effects 0.000 abstract description 2
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- 238000009423 ventilation Methods 0.000 abstract 1
- 238000009395 breeding Methods 0.000 description 5
- 230000001488 breeding effect Effects 0.000 description 5
- 238000005286 illumination Methods 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 4
- 239000007789 gas Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 230000006978 adaptation Effects 0.000 description 2
- 235000004879 dioscorea Nutrition 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 239000010977 jade Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
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- 229920001817 Agar Polymers 0.000 description 1
- 241000255789 Bombyx mori Species 0.000 description 1
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- 240000001811 Dioscorea oppositifolia Species 0.000 description 1
- 235000003416 Dioscorea oppositifolia Nutrition 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000010413 gardening Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000012567 medical material Substances 0.000 description 1
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- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
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- 229910052698 phosphorus Inorganic materials 0.000 description 1
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Abstract
The invention discloses a method for culturing a yam seed tuber by using a yam tissue culture seedling, and belongs to the technical field of agricultural production. The yam seedling root system tissue culture seedling is cultured through shading treatment at the bottom of a tissue culture bottle; the seedling root system tissue culture seedling is hardened and transplanted; local light transmission water saving coverage is combined with ventilation at a transplanting stage, so the yam tissue culture seedling can adapt to quick change of external environment after transplantation, and the transplantation survival rate reaches 71.4 percent; through a series of steps such as strict seedbed management, the finally obtained proportion of the tissue culture seed tuber of more than 5g is 46.3 percent. By using the tissue culture seedling for propagation of the yam seed tubers, industrialized production of the yam seed tuber propagation can be realized. The method has the characteristics of high propagation coefficient, high propagation speed and less viruses of the seed tubers.
Description
Technical field
The present invention relates to a kind of method utilizing Rhizoma Dioscoreae tissue cultured seedling to cultivate Rhizoma Dioscoreae potato seed, belong to field of agricultural production technologies.
Background technology
Rhizoma Dioscoreae (Dioscorea Opposita Thunb.), plants for Dioscoreaceae Wild yam perennial herbaceous stem prehensile liana
Thing, all there is plantation various places, China north and south.Because it has higher nutritive value and medical value, it is medical material and the biography in China tunnel
The states such as system exporting, its product situation of selling well Southeast Asia and Japan.Rhizoma Dioscoreae generally uses tuber to breed, and breeding coefficient is low, and
Production cost is high, and utilizes tuber breeding to make virus constantly accumulation cause kind of a sexual involution for a long time, has had a strong impact on quality and product
The raising of amount.
Tissue culture is possible not only to improve breeding coefficient, and utilizes detoxication and tissue culture technology can effectively keep kind kind
Property, improving quality, improves yield.But test tube Seedling exists domestication to be required strict, and transplanting survival rate is on the low side, potato seed yield is on the low side etc. is asked
Topic.Especially under the general condition of culture of Rhizoma Dioscoreae tissue cultured seedling, generate aerial root, to external world the adaptation ability of environment and resistance more
Difference, transplanting can cause seedling mortality, and therefore Rhizoma Dioscoreae tissue cultured seedling transplanting survival rate always limits the application of Rhizoma Dioscoreae tissue culture expanding propagation
Basic reason.Substrate, in addition to having the function of fixing crop root, it is often more important that for root system create good water,
The conditions such as fertilizer, gas.The matrix species impact on Rhizoma Dioscoreae tissue cultured seedling transplanting survival rate of existing source investigation, such as document 1: Tang
Rainbow, Wu Jiali, Zhang Ling, Chen Zilin, different substrates is on oxtail Rhizoma Dioscoreae tissue cultured seedling potato block quality and the impact of growth, China's gardening literary composition
Pluck, 2010, (9): 26-31.Also have part information that Rhizoma Dioscoreae tissue cultured seedling is transplanted to summarize, such as document 2: Wang Guolian, Chen Ming, grandson
Yu Dong, Zhong Xiujuan, the Shoot Tip Culture of Rhizoma dioscoreae and Fast-propagation, Jiangsu's agriculture science, 2003, (3): 77-78；Document 3: Zhou Yu
The tinkling of pieces of jade, Yu Huilin, Sun Fengling, Jiang Shuguang, Li Xinyu, Liu Guangqing, Zhang Fujuan, the tissue culture of favour building Rhizoma Dioscoreae grinds with fast breeding technique
Study carefully, Agriculture of Anhui science, 2009, (20): 9407-9408.
These are reported as Rhizoma Dioscoreae tissue cultured seedling transplantation technique and have established certain basis, but Rhizoma Dioscoreae tissue cultured seedling is transplanted the most in time
Relatively high-survival rate can be obtained, how to carry out scientific management after transplanting and then obtain a large amount of original silkworm egg and define still without clear and definite.
This research is from the root system of Rhizoma Dioscoreae tissue cultured seedling and the measure such as seedbed management after transplanting seedbed preparation, transplanting time, method for transplanting, transplanting
Set about, fundamentally solve Rhizoma Dioscoreae tissue cultured seedling and transplant the problem difficult, survival rate is low, it is provided that cultivate Rhizoma Dioscoreae with Rhizoma Dioscoreae tissue cultured seedling
Group cultivates the effective ways of potato.
Summary of the invention
Technical problem
Present invention mainly solves technical problem is that: increases substantially the method for transplanting of Rhizoma Dioscoreae group training transplant survival, utilizes
Rhizoma Dioscoreae tissue cultured seedling carries out potato seed seedbed and concentrates the technical method bred.
Technical scheme
For solving the problems referred to above, a kind of method utilizing Rhizoma Dioscoreae tissue cultured seedling to cultivate Rhizoma Dioscoreae potato seed of the present invention, the technology of employing
Scheme is:
(1) root that grows directly from seeds is cultivated: utilize 1/2MS+6-BA0.1mg L-1+NAA2.0mg·L-1Root media, uses
Shading treatment bottom tissue culture bottle, it is ensured that not irradiation bottom tissue cultured seedling, carries out root culture to Rhizoma Dioscoreae tissue cultured seedling.Cultivation temperature (25
± 2) DEG C, light application time 12h d-1, cultivation cycle is 60～70d, i.e. obtains the Rhizoma Dioscoreae group training test tube Seedling that root is master that grows directly from seeds；
(2) locally water conservation covers and transplants: the test tube Seedling of step (1) grows to high 4～6, and grow directly from seeds root 10～when 15, through refining
After Seedling processes, it is transplanted to substrate seedbed；After transplanting, it is placed on complete for tissue cultured seedling aerial parts on tissue cultured seedling with light transmission container is outstanding gently
Full clad, carries out local water conservation and covers tissue cultured seedling, seedbed covered with sunshade net；In transplanting 3d, whole day keeps covering and sunshade；Move
Take tissue culture bottle and sunshade net off at 5 in afternoon～the next morning 8 after planting 3d, after 8, again cover tissue culture bottle water conservation and screening
Screened postive is sheltered from heat or light, every 1d repetitive operation 1 time；Take tissue culture bottle completely off when transplanting 20d, after 60d, take sunshade net completely off.
Described local water conservation covers in method for transplanting, and tissue cultured seedling, after seedling exercising processes, is taken out of gently by test tube Seedling with tweezers,
With clear water, root surface culture medium is cleaned, plant in seedbed.
Described seedling exercising is: test tube Seedling grows to high 4～6, grow directly from seeds root 10～15, and during average root length 4～6cm, whole bottle moves
To with culturing room temperature difference 5 DEG C within outdoor, after placing 5～6d under scattered light, open bottle cap 1d.
Described seedbed is: seedbed uses sufficient fertilizer, ploughs deeply thin rake, the front 5～7cm degree of depth of being dug in seedbed of transplanting, seedbed width 1m,
Being smashed by soil and cover Nutrition Soil 4～5cm, uniformly sprinkling 4.5% effective cypermethrin is around seedbed and seedbed, waters foot water standby；
Seedbed water management after transfer: tissue cultured seedling is transplanted to water and determined root water the same day, waters a water every 2～3d later；Tissue cultured seedling survives
After to water in time, seedbed should be seen wet；Fertilizer management: expanding stage imposes 45% common NPK 0.15kg m2；Seedling
Tortuous bow is taken for shoot growth on Chuan.
Beneficial effect
1) present invention cultivates the root that grows directly from seeds is the suction of nutrient substance after main Rhizoma Dioscoreae tissue cultured seedling, beneficially Rhizoma Dioscoreae tissue cultured seedling are transplanted
Receive and Fa Xingen, lay the foundation for improving transplanting survival rate.
(2) present invention local water conservation covers method for transplanting, utilizes the transparent water conservation container of suitable size to carry out local water conservation
Cover, in conjunction with practicing Seedling, sunshade and the management technique of progressive adaptation, coordinate Rhizoma Dioscoreae tissue cultured seedling root of hair and keep the aqueous vapor relation of blade
Etc. environmental condition, Rhizoma Dioscoreae tissue cultured seedling gradually adapts to huge environmental change, makes Rhizoma Dioscoreae tissue cultured seedling transplanting survival rate bring up to 70%
Above.
(3) the supporting application of the measures such as the present invention was transplanted by optimum period, seedbed precision management, enables the tissue cultured seedling survived
Form bigger potato seed.By transplanting and precision management in good time, it is ensured that the tissue cultured seedling after transplant survival is formed then and meets potato seed
The Rhizoma Dioscoreae tuber required.Obtaining more than 5g group and cultivating potato ratio is 46.3%.
Accompanying drawing explanation
Fig. 1 yam test tube seedling rooting type: A: aerial root；B: grow directly from seeds root, root length；C: grow directly from seeds root, root is short
Fig. 2 yam test tube transplantation of seedlings: A: aerial root type group seedlings cultivating is transplanted；B: root length of growing directly from seeds tissue cultured seedling is transplanted；C: grow directly from seeds
The short tissue cultured seedling of root is transplanted
The group of Fig. 3 results cultivates potato
Detailed description of the invention
The present invention is further illustrated below by based on " one utilizes Rhizoma Dioscoreae tissue cultured seedling to cultivate potato seed method ".
1 materials and methods
1.1 materials: Taizhou Rhizoma Dioscoreae test tube Seedling
1.2 experimental working technique
1. tissue cultured seedling root culture: root media is 1/2MS+6-BA0.1mg L-1+NAA2.0mg·L-1, high temperature is high
Pressure sterilizing.By Taizhou Rhizoma Dioscoreae test tube Seedling (Han Xiaoyong, Yan Ruixia, Yin Jianmei, Zhang Peitong, Guo Wen with 1 or more than 1 leaf
Fine jade, Li Chunhong, Taizhou Rhizoma Dioscoreae tissue culture quick breeding technical research, Zhejiang Agriculture journal, 2014,26 (2): 344-347) stem section
Cut, proceed to root media and cultivate, medium agar concentration 6.5g L-1, sucrose 30g L-1, pH value 5.8, cultivation temperature
(25 ± 2) DEG C, light application time 12h d-1.Using bottom tissue cultured seedling not irradiation, bottom tissue cultured seedling, illumination is about 600lx, tissue cultured seedling
Bottom illumination is that tri-kinds of training methods of 2500lx are cultivated, and cultivation cycle is 60～70d.
2. seedling exercising: test tube Seedling grows to high 4～6, grow directly from seeds root 10～15, during average root length 4～6cm, whole bottle move on to
Outdoor within culturing room's temperature difference 5 DEG C, places 5～6d under scattered light, opens bottle cap 1d.
3. seedbed prepares: seedbed uses sufficient fertilizer (N+P2O5+K2O >=7%, organic >=30%, Nanjing Changhong fertilizer is public
Department) 1.5kg m2, plough deeply thin rake, before transplanting, seedbed dug spade degree of depth soil (5～7cm), seedbed width 1m, soil is smashed and covers battalion
(organic culture substrate, the content of organic matter 50% in substrate, nitrogen, phosphorus, potassium content 2.5%, Zhenjiang emerging agriculture fertilizer has to support soil
Limit company) 4～5cm, uniformly sprinkling 4.5% effective cypermethrin is around seedbed and seedbed, waters foot water standby.
4. local water conservation covers method for transplanting: Rhizoma Dioscoreae tissue cultured seedling should be in May 20 (southern area of Jiangsu Province) front transplanting.Treat tissue cultured seedling
Grow to high 4～6, grow directly from seeds root 10～15, during average root length 4～6cm, after seedling exercising processes, it is transplanted to substrate seedbed.
Locally water conservation covering method for transplanting is: after training tissue culture seedling, is taken out of gently by tissue cultured seedling with tweezers, accomplishes not as far as possible
Injure root system, with clear water, root surface culture medium is cleaned, plant in preprepared seedbed, transplant to water the same day and determine root water, after
A water is watered every 2～3d.
After transplanting, maybe can be placed on tissue cultured seedling by outstanding gently for the light transmission container of complete for tissue cultured seedling aerial parts clad with tissue culture bottle
Top, carries out local water conservation and covers tissue cultured seedling, seedbed covered with sunshade net；
In transplanting 3d, whole day keeps covering and sunshade；
Take covering light transmission container and sunshade net off at 5 in afternoon～the next morning 8 after 3d, again cover thoroughly after 8
Light container water conservation and covered with sunshade net shelter from heat or light, every 1d repetitive operation 1 time；
Take light transmission container completely off when transplanting 20d, after 60d, take sunshade net completely off.
5. seedbed management: water management: tissue cultured seedling is transplanted to water and determined root water the same day, waters a water every 2～3d later；Group
Seedlings cultivating to water after surviving in time, and seedbed should be seen wet；Fertilizer management: expanding stage imposes 45% common NPK (N15-
P15-K15, total nutrient >=45%, Jurong Aurion specialization fertilizer company limited) 0.15kg m2；Routine Management: take tortuous bow on seedbed
For shoot growth.
2, result of the test
2.1 tissue cultured seedling root culture
As can be seen from Table 1, bottom intensity of illumination has influence on test tube Seedling aerial root rate.Bottom not irradiation, test tube Seedling gas is raw
Root rate is minimum, is 5.4%, and root system predominantly grows directly from seeds root.This is likely due to bottom test tube Seedling not irradiation, can be big with simulating plant
Field growing environment, thus reduce the generation of aerial root.
The different training method impact on tissue cultured seedling aerial root rate of table 1
| Training method | Aerial root rate (%) |
| Bottom not irradiation | 5.4 |
| Bottom illumination about 600lx | 44.4 |
| Bottom illumination is 2500lx | 93.2 |
2.2 transplant
Being divided three classes by Rhizoma Dioscoreae tissue cultured seedling before Yi Zaiing: type I: height of seedling 4～6, root system is mainly aerial root (aerial root rate
More than 90%)；Type II: height of seedling 4～6, root system mainly grows directly from seeds root (aerial root rate be less than 10%), and grow directly from seeds root 10～15
Root, average root length 4～6cm；Type III: height of seedling 4～6, root system mainly grows directly from seeds root (aerial root rate be less than 10%), average root
Long 2～3cm.
By the inventive method, three class tissue cultured seedlinies being carried out local water conservation and covers transplanting, result shows (table 2): after transplanting 30d,
Type I, II, III test tube shoot survival percent is respectively 43.6%, 71.4% and 68.2%, and this shows the Rhizoma Dioscoreae group training that root is master of growing directly from seeds
Transplantation of seedlings survival rate is high, and real root length is the longest more strong, and survival rate is the highest.And do not take the type of local water conservation covering treatment
I test tube shoot survival percent is only 3.3%, and illustrating that the local water conservation of the present invention covers transplantation technique is to determine Rhizoma Dioscoreae tissue cultured seedling survival rate
Main factor, and whether tissue cultured seedling root system is the root that grows directly from seeds also has a significant effect the survival rate of tissue cultured seedling.
The impact on tissue cultured seedling survival rate of table 2 method for transplanting
2.3 potato seed yield
To the method test community of the present invention, (tissue cultured seedling Type II: height of seedling 4～6, root system mainly grows directly from seeds root, and gas is raw
Root rate is less than 15%, grow directly from seeds root 10～15, average root length 4～6cm) group gathered in the crops cultivates potato and carries out adding up (table 3), transplant into
Tissue cultured seedling 106 strain alive, gathers in the crops potato seed 122.Potato seed is carried out classification: I grade (more than 5g), II grade (1～5g), III grade (1g with
Under), wherein obtaining more than 5g group and cultivating potato ratio is 46.3% (table 1).
Potato classification cultivated by 3 groups of table
| I grade of potato seed ratio/% | II grade of potato seed ratio/% | III grade of potato seed ratio/% |
| 46.3 | 31.7 | 22 |
When being understood by, to those skilled in the art, can modifications and variations the most in addition,
And all these modifications and variations all should belong to the protection domain of claims of the present invention.
Claims (1)
1. one kind utilizes the method that Rhizoma Dioscoreae tissue cultured seedling cultivates Rhizoma Dioscoreae potato seed, it is characterised in that including:
(1) root that grows directly from seeds is cultivated: utilize 1/2MS+6-BA0.1mg L-1+ NAA2.0mg·L-1Root media, employing group
Shading treatment bottom training bottle, it is ensured that not irradiation bottom tissue cultured seedling, carries out root culture to Rhizoma Dioscoreae tissue cultured seedling, cultivation temperature (25 ±
2) DEG C, light application time 12h d-1, cultivation cycle is 60～70d, i.e. obtains the Rhizoma Dioscoreae group training test tube Seedling that root is master that grows directly from seeds；
(2) locally water conservation covers and transplants: the test tube Seedling of step (1) grows to height of seedling 4～6, and grow directly from seeds root 10～when 15, through seedling exercising
After process, it is transplanted to substrate seedbed；After transplanting, it is placed on complete for tissue cultured seedling aerial parts on tissue cultured seedling with light transmission container is outstanding gently
Clad, carries out local water conservation and covers tissue cultured seedling, seedbed covered with sunshade net；In transplanting 3d, whole day keeps covering and sunshade；Transplant
Take tissue culture bottle and sunshade net off at 5 in afternoon～the next morning 8 after 3d, after 8, again cover tissue culture bottle water conservation and sunshade
Net shelters from heat or light, every 1d repetitive operation 1 time；Take tissue culture bottle completely off when transplanting 20d, after 60d, take sunshade net completely off；
Described local water conservation covers in method for transplanting, and tissue cultured seedling, after seedling exercising processes, is taken out of gently by test tube Seedling with tweezers, with clear
Root surface culture medium is cleaned by water, plants in seedbed；
Described seedling exercising is: test tube Seedling grows to high 4～6, grow directly from seeds root 10～15, during average root length 4～6cm, whole bottle move on to
Outdoor within culturing room's temperature difference 5 DEG C, after placing 5～6d, opens bottle cap 1d under scattered light；
Described seedbed is: seedbed uses sufficient fertilizer, ploughs deeply thin rake, before transplanting, seedbed is dug the 5～7cm degree of depth, and seedbed width 1m, by soil
Smashing and cover Nutrition Soil 4～5cm, uniformly sprinkling 4.5% effective cypermethrin is around seedbed and seedbed, waters foot water standby；
Seedbed water management after transplanting: tissue cultured seedling is transplanted to water and determined root water the same day, waters a water every 2～3d later；Tissue cultured seedling
To water in time after surviving, seedbed should be seen wet；Fertilizer management: expanding stage imposes 45% common NPK 0.15kg m2；
Tortuous bow is taken for shoot growth on seedbed.
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| CN107155843B (en) * | 2017-05-16 | 2020-05-08 | 江苏省农业科学院 | Improved method for industrially transplanting tissue culture seedlings of Chinese yams and cultivating potato seeds |
| CN111280037A (en) * | 2020-02-25 | 2020-06-16 | 江苏省农业科学院 | Water culture propagation and seed potato preservation method for ginseng and potato Chinese yams |
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