CN109744142A - A kind of santal tissue-culturing quick-propagation method for culturing seedlings - Google Patents
A kind of santal tissue-culturing quick-propagation method for culturing seedlings Download PDFInfo
- Publication number
- CN109744142A CN109744142A CN201710718937.6A CN201710718937A CN109744142A CN 109744142 A CN109744142 A CN 109744142A CN 201710718937 A CN201710718937 A CN 201710718937A CN 109744142 A CN109744142 A CN 109744142A
- Authority
- CN
- China
- Prior art keywords
- culture
- santal
- days
- tree
- micro
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Cultivation Of Plants (AREA)
Abstract
The present invention provides a kind of santal tissue-culturing rapid propagation micro cuttage method for culturing seedlings, include the following steps, the culture of S1, rudiment bar；S2, explant disinfection；S3, clump bud induction；S4, squamous subculture；S5, pre- culture of rootage；S6, hardening culture；S7, micro cuttage transplanting.The present invention is using the select tree of the 1/3 provenance test woods more than period of felling in turn as culture materials, based on tissue-culturing rapid propagation, bottle seedling of taking root in advance is point of penetration, by adjusting shoot proliferation, hormone concentration and proportion, sucrose concentration, transplant the series techniques method such as Jie's base and proportion, moisture control and nutrient supplement, Disease management, the present invention provides a set of santal micro cuttages to produce matching technology, solve the problems, such as that santal culture of rootage is not taken root and fall leaves death, realize pre- culture of rootage after 45 days, 98% bottle seedling height reaches 3.5cm or more, and robust growth is not fallen leaves；Realize that micro cuttage rooting rate of taking root in advance reaches 60% or more；Realize the large-scale production of clonal tissue culture nursery stock.
Description
Technical field
The present invention relates to the fine-variety breeding of tropical forest and seedling propagation technical fields, organize more particularly, to a kind of santal
Micro cuttage method for culturing seedlings is quickly bred in culture.
Background technique
Santal (Santalum album L) is a kind of evergreen semiparasite little Qiao that Santalaceae (Santalaceae) santal belongs to
Wood originates in Indonesia, it be it is a kind of integrate medicinal, Buddhism articles, fragrance, hand work material for carving on it is important
Economic tree.
The history of existing more than 1000 years of consumption of santal, from ancient times universally acknowledged is luxury goods.Currently, santal provides both at home and abroad
Source is sharply being reduced, and whole world annual output is less than 1000 tons now, and sandalwood fragrance underproduces 1 ton, and seriously supply falls short of demand in market.Cause
This, santal planting industry becomes newborn rising industry, has the very big prospect of marketing.Currently, China Guangdong, Guangxi, sea
Sandalwood cultivation is widelyd popularize in the provinces and regions such as south, Yunnan, Sichuan, Fujian, and obtains the heartwood with economic value, but lacks high-quality
Santal nursery stock, more shortage santal choiceness, limit the development of santal planting industry industry.Excellent maternal plant is selected, is carried out
Clonal reproduction is the key technology for realizing Sandalwood cultivation improved variety and clone, captures santal tissue culture nursery stock and takes root difficulty, raw
Root culture is not taken root and falls leaves death, the low problem of transplanting success, is that santal breeding and cultivation must solve key technology hardly possible
Topic.
Original technology situation: tissue cultures are transplanted from explant disinfection → Multiplying culture → culture of rootage → nursery.It utilizes
The above method carries out numerous fastly have the following deficiencies: to santal
Santal is semiparasitic plant, and after embryo germination soon, root system can form sucker, be adsorbed on other root systems of plant
On, absorb moisture and nutrition from the root system of other plants, this characteristic determines that santal tissue culture and inducement is taken root difficulty, or induction
After forming root restriction and short, it is difficult to continue development and form the good root system that can support transplanting survival.Therefore, conventional tissue culture
Culture of rootage difficulty is big, and transplanting survival difficulty is also big.In addition, santal root system nutrition absorbability is weak, Tube plantlets migration process
It is also the technical problem for needing especially to solve that nutrition, which how is replenished in time,.
Summary of the invention
The main purpose of the present invention is to provide a kind of santal tissue-culturing rapid propagation micro cuttage method for culturing seedlings.This method aims to solve the problem that
Difficult, culture of rootage that santal tissue cultures are taken root is not taken root and fall leaves problem dead, that transplanting success is low, realization field micro cuttage
Success, achievees the purpose that large-scale production excellent strain nursery stock.Its major technique is the research method using tissue rapid propagation, is realized
The clone sprout of the more than period of felling in turn high-quality santal maternal plant of biological control 1/3 passes through special pre- rooting treatment, culture growth
Stalwartness is not fallen leaves, highly in the healthy sprout of 3.5cm or more, in conjunction with cuttage technique, organically tissue cultures and cutting propagation
Combine, realize the transplanting of micro cuttage formula, transplants rooting rate up to 60% or more.
Method is as follows: a kind of santal tissue-culturing rapid propagation micro cuttage method for culturing seedlings includes the following steps:
The culture of S1, rudiment bar: in autumn and winter, high according to tree in the fine provenance woods that the age of tree was more than 1/3 period of felling in turn,
The indexs such as the diameter of a cross-section of a tree trunk 1.3 meters above the ground and symbiosis tree quantity choose select tree, select illumination condition on select tree good, the branch closest to ground of robust growth
Item carries out lopping cutting handle, and lopping retains 40~60cm length, removes the former coppice shoot in lopping, and spring next year germination rate is reachable
95%；
S2, explant disinfection: in spring and summer, the select tree coppice shoot for 20~40cm that acquisition lopping is sprouted is wiped out blade and is stayed
Petiole, 1~for 24 hours in 10min cleaned using ultrasonic drilling machine clear water, then on super quiet workbench, with the mercury disinfection 13 of 0.1g/L~
20min sterilizes success rate up to 60% or more；
S3, clump bud induction: the explant after disinfection accesses induction clump bud in clump bud inducement cultivation base and sprouts, medium pH
Value is 5.8~6.2, the element of the MS including improvement 1, content are as follows: NH4NO3350~550mg/L, KH2PO4170~
255mg/L、MgSO4.7H2370~550mg/L of O, KNO3 1900mg/L、CaCl2 400mg/L、Ca(NO3)2.4H2O 750mg/
L, (NH4)2SO4260mg/L, microelement 1.5MS, organic matter 1.0MS, 0.10~0.2mg/L of hormone 6BA, IBA 0.2mg/
L, 30~35mg/L of sucrose, squamous subculture condition be intensity of illumination be 2000~3000LX, light application time is 10 hours/day, training
Supporting temperature is 22~28 DEG C.
S4, squamous subculture: after clump bud derives, it is transferred to santal subculture multiplication medium, condition of culture removes hormone concentration
Change it is outer, other conditions with step S3, breeding rate up to 2.5 times or more, expand it is numerous based on the squamous subculture period be 30~35
It, the squamous subculture period that seedling is produced while expanding numerous is 40~50 days.The hormone of subculture medium changes are as follows: and 6BA0.05~
0.10~0.15mg/L of 0.15mg/L, IBA；
S5, pre- culture of rootage: choosing height in squamous subculture, in 2.5cm or more, the sprout access that blade is sufficiently spread out is pre-
Root media carries out pre- culture of rootage and is transferred to hardening in hardening greenhouse after culture 25~35 days；
Pre- culture of rootage is the element of the MS of improvement 2, content are as follows: NH4NO3560~1320mg/L, KH2PO4256~
340mg/L、MgSO4 7H2O 148mg/L、KNO3760~1520mg/L, CaCl2 130mg/L、Ca(NO3)2 4H2O
750mg/L, microelement 1MS, organic matter 1MS, hormone GGR 0.4mg/L, IBA 0.2mg/L, NAA 0.2mg/L, sucrose 40
~50mg/L.
S6, hardening culture: the time of pre- culture of rootage is different because of season, is after 25~35 days, then, without culture of transferring
Base is directly transferred to hardening in the hardening greenhouse that intensity of illumination is 5000~10000Lx, and the hardening time is 15~30 days.Work as sprout
Highly >=4cm or more when, in time transplant, reduce practice seedling overlong time, pollution rate increase, reduce bottle seedling bottle outlet rate；
S7, micro cuttage transplanting: it is suitable for the matrix formulations of micro cuttage transplanting are as follows: vermiculite: peat soil: the volume ratio of yellow soil
For 8:1:1.Matrix is sterilized with 0.5% potassium permanganate before micro cuttage, the different fungicide of use per week carry out disease after micro cuttage
Prevention and treatment.Because santal Tube plantlets micro cuttage rootage duration is long, required nutrient cannot be replenished in time in conventional fertilizer application.Pre- sprout of taking root
After transplanting 20 days, a great number of elements solution of 1 1/4MS~1/3MS of foliage-spray per week supplements nutrient.After transplanting 23 days, often
10 days foliage-sprays, 1 1.5~2.0MS trace element solution.After the transplanting of santal Tube plantlets micro cuttage, early period should be noted that moisturizing
And foliar spray, and cover sunshade net reduction intensity of illumination, then gradually decrease spray timings and reinforce illumination.Transplanting 2~2.5
After a month, rooting rate is transplanted up to 60% or more.
Preferably, in step S1, the above selected select tree of life in 8 years, the branch lopping closest to ground retains 40~60cm long
Degree, 95% or more spring next year germination rate.
Preferably, in step S2, explant disinfection: the coppice shoot in acquisition select tree lopping, 1~interior for 24 hours clear using ultrasonic drilling machine
It washes, sterilizes 13~20min with the mercury of 1mg/L, sterilize success rate up to 60% or more.
Preferably, shoot proliferation multiple step S3, in S4 reaches 2.5 times or more.Condition of culture is intensity of illumination 2000
~3000LX, 10 hour/day of light application time, 22~28 DEG C of cultivation temperature.The culture medium constituent content of improvement 1 are as follows: NH4NO3 350
~550mg/L, KH2PO4170~255mg/L, MgSO4.7H2O.370~550mg/L, KNO3 1900mg/L、CaCl2
400mg/L、Ca(NO3)2.4H2O 750mg/L, (NH4)2SO4260mg/L, microelement 1.5MS, organic matter 1.0MS, hormone
0.05~0.2mg/L of 6BA, IBA 0.2mg/L, 30~35mg/L of sucrose；
Preferably, in step s 5,2 pre- root medias of improvement include the element of following content, NH4NO3560~
1320mg/L、KH2PO4256~340mg/L, MgSO4.7H2O 148mg/L、KNO3760~1520mg/L, CaCl2 130mg/
L、Ca(NO3)2 4H2O 750mg/L, microelement 1.0MS, organic matter 1.0MS, hormone GGR 0.4mg/L, IBA 0.2mg/L,
NAA 0.2mg/L, 40~45mg/L of sucrose.
Preferably, step S6, in S7, vermiculite: peat soil: the matrix that yellow soil volume ratio is 8:1:1 is that micro cuttage is best
Matrix.After transplanting 20 days, 1/4~1/3MS a great number of elements of foliage-spray per week, 1.5~2.0MS microelement are supplementary fertilizer
Best mode, 2 after half a month, transplants rooting rate up to 60% or more.
Effect and advantage of the present invention:
1, it uses 1/3 period of felling in turn of the age of tree above santal select tree for propagation material, santal choiceness nursery stock can be bred,
To realize the clone and improved variety of Sandalwood cultivation, the increment and timber product of santal artificial forest can be finally increased substantially
Matter increases substantially the economic benefit of Sandalwood cultivation.
2, by the induction of specific clump bud, the development of three culture mediums of squamous subculture and pre- root media, Gao Fan is realized
The pre- bottle seedling of taking root of the proliferation times and high quality grown reduces production cost, improves productivity effect.
3, pass through specific micro cuttage grafting matrix: vermiculite: peat soil: yellow soil=8:1:1, specific nutrient supplement,
The series techniques measures such as miscellaneous bacteria control, moisture control realize that micro cuttage transplants rooting rate up to 60% or more.
4, the present invention realizes that tissue cultures and cutting propagation organically combine, and breaks through santal culture of rootage and does not take root or take root
Difficult bottleneck, accomplishes scale production.
5, operation of the present invention is simple, at low cost, with short production cycle.
Specific embodiment
It is explained further the present invention with reference to embodiments, but embodiment does not do any type of limit to the present invention
It is fixed.
Embodiment 1: a kind of santal tissue-culturing rapid propagation micro cuttage method for culturing seedlings includes the following steps:
Exploiting entity: this laboratory santal culture materials is taken a little: Zhanjiang
The culture of S1, rudiment bar: in 2010 autumns, 3 plants of select trees for selecting life in 8 years above select illumination condition good, growth
Healthy and strong branch lopping immediate from the ground retains 40~60cm length, and the former coppice shoot in peace and quiet lopping.
S2, explant disinfection: next year takes the select tree coppice shoot of 20~40cm, wipes out blade and stays petiole, uses ultrasonic wave in 12h
Machine cleans 10min, and on super quiet workbench, sterilizes 13~20min with the mercury of 0.1mg/L, sterilizes success rate up to 60% or more.
S3, clump bud induction: the explant after disinfection accesses in clump bud inducement cultivation base, and Medium's PH Value is 5.8~6.2,
The element of MS including improvement 1, content are as follows: NH4NO3350~550mg/L, KH2PO4170~255mg/L, MgSO4。
7H2370~550mg/L of O, KNO3 1900mg/L、CaCl2 400mg/L、Ca(NO3)24H2O 750mg/L, (NH4)2SO4
260mg/L, microelement 1.5MS, organic matter 1.0MS, 0.10~0.2mg/L of hormone 6BA, IBA 0.2mg/L, sucrose 30~
35mg/L；Squamous subculture condition be intensity of illumination be 2000~3000Lx, 10 hour/day of light application time, cultivation temperature 22~28
℃。
S4, squamous subculture: after clump bud derives, it is transferred to santal subculture multiplication medium, condition of culture removes hormone concentration
Change it is outer, other conditions with step S3, expand it is numerous based on the squamous subculture period be 30 days, produced while expanding numerous seedling after being commissioned to train
Supporting the period is 45 days, and the hormone of subculture medium changes are as follows: hormone 6BA0.05~0.15mg/L, IBA0.10~0.15mg/L；
The same S3 of condition of culture
S5, pre- culture of rootage: choosing height in squamous subculture, in 2.5cm or more, the sprout access that blade is sufficiently spread out is pre-
Root media carries out pre- culture of rootage and is transferred to hardening in hardening greenhouse after culture 25~35 days, improves the 2 pre- life of MS element
Root culture medium, the element comprising following content: NH4NO3560~1320mg/L, KH2PO4256~340mg/L, MgSO4
7H2O 148mg/L、KNO3760~1520mg/L, CaCl2 130mg/L、Ca(NO3)2 4H2O 750mg/L, microelement
1MS, organic matter 1MS, hormone GGR 0.4mg/L, IBA 0.2mg/L, NAA0.2mg/L, 40~50mg/L sucrose.
S6, hardening culture: after taking root sprout culture 25~35 days in advance, it is transferred to the refining that intensity of illumination is 5000~10000LX
Hardening in seedling greenhouse, the hardening time is 15~30 days, as sprout height >=4cm or more, transplants in time, reduces and practice seedling time mistake
Long, pollution rate increases, and reduces bottle seedling bottle outlet rate；
S7, micro cuttage transplanting: use vermiculite: peat soil: yellow soil volume ratio is the micro cuttage matrix of 8:1:1, micro cuttage
It is preceding to sterilize matrix with 0.5% potassium permanganate.3~5 days full lid films after the transplanting of the sprout of pre- culture of rootage and hardening, and hide
The sunshade net that two layers 75% of lid, it is daily to open 4 times by spraying；5th~10 day opening film retains sunshade net, and auto spraying 6 times/
Day, 10 seconds every time；The sunshade net of one layer 75% of reservation in 11st~20 day, 5 times/day of auto spraying, every time 10 seconds；It is automatic after 20 days
It is 4 times/day, every time 10 seconds spraying.It is per week after transplanting to carry out disease control using 1/1000 different fungicide.Transplanting 20 days
Afterwards, nutrient is supplemented using 1/3MS a great number of elements, supplement per week is primary later.Transplanting 23 days after, supplement 2MS moderate-element and
Microelement, supplement is primary within later 10 days.Transplanting 2 after half a month, transplants rooting rate up to 60% or more.
Embodiment 2: a kind of santal tissue-culturing rapid propagation micro cuttage method for culturing seedlings includes the following steps:
Exploiting entity: this laboratory cultures material is taken a little: the village Jiao Tang, the county Gao Yao
The culture of S1, rudiment bar: in October, 2015 selects 4 plants of select trees from the 8 years non-hibernating eggs source woodss in the pool Gao Yaojiao village, selection
Illumination condition is good, the branch lopping immediate from the ground of robust growth, retains 40~60cm length, and the original in peace and quiet lopping
Coppice shoot.
S2, explant disinfection: in July, 2016 takes the select tree coppice shoot of 20~40cm, wipes out blade and stays petiole, uses in 12h
Ultrasonic drilling machine cleans 10min, and on super quiet workbench, sterilizes 13~20min with the mercury of 0.1mg/L, disinfection success rate reaches
60% or more.
S3, clump bud induction: the explant after disinfection accesses in clump bud inducement cultivation base, and Medium's PH Value is 5.8~6.2,
The element of MS including improvement 1, content are as follows: NH4NO3350~550mg/L, KH2PO4170~255mg/L, MgSO47H2O
370~550mg/L, KNO3 1900mg/L、CaCl2 400mg/L、Ca(NO3)24H2O 750mg/L, (NH4)2SO4 260mg/L、
Microelement 1.5MS, organic matter 1.0MS, 0.10~0.2mg/L of hormone 6BA, IBA 0.2mg/L, 30~35mg/L of sucrose；
Squamous subculture condition be intensity of illumination be 2000~3000Lx, 10 hour/day of light application time, cultivation temperature 22~28
℃。
S4, squamous subculture: after clump bud derives, it is transferred to santal subculture multiplication medium, condition of culture removes hormone concentration
Change it is outer, other conditions with step S3, expand it is numerous based on the squamous subculture period be 30 days, produced while expanding numerous seedling after being commissioned to train
Supporting the period is 50 days, and the hormone of subculture medium changes are as follows: hormone 6BA0.05~0.15mg/L, IBA0.10~0.15mg/L；
The same S3 of condition of culture
S5, pre- culture of rootage: choosing height in squamous subculture, in 2.5cm or more, the sprout access that blade is sufficiently spread out is pre-
Root media carries out pre- culture of rootage and is transferred to hardening in hardening canopy after culture 25~35 days, improves 2 MS element and takes root in advance
Culture medium, the element comprising following content: NH4NO3560~1320mg/L, KH2PO4256~340mg/L, MgSO4
7H2O148mg/L、KNO3760~1520mg/L, CaCl2 130mg/L、Ca(NO3)2 4H2O 750mg/L, microelement 1MS,
Organic matter 1MS, hormone GGR 0.4mg/L, IBA 0.2mg/L, NAA0.2mg/L, 40~50mg/L sucrose.
S6, hardening culture: after taking root sprout culture 25~35 days in advance, it is transferred to the refining that intensity of illumination is 5000~10000LX
Hardening in seedling greenhouse, the hardening time is 15~30 days, as sprout height >=4cm or more, transplants in time, reduces and practice seedling time mistake
Long, pollution rate increases, and reduces bottle seedling bottle outlet rate；
S7, micro cuttage transplanting: use vermiculite: peat soil: yellow soil volume ratio is the micro cuttage matrix of 8:1:1, micro cuttage
It is preceding to sterilize matrix with 0.5% potassium permanganate.0~4 day full lid film after the transplanting of the sprout of pre- culture of rootage and hardening, and hide
The sunshade net that two layers 75% of lid, it is daily to open 4 times by spraying；5th~10 day opening film retains sunshade net, and auto spraying 6 times/
Day, 10 seconds every time；The sunshade net of one layer 75% of reservation in 11st~20 day, 5 times/day of auto spraying, every time 10 seconds；It is automatic after 20 days
It is 4 times/day, every time 10 seconds spraying.It is per week to carry out disease control using 1/1000 different fungicide.After transplanting 20 days, use
1/4MS a great number of elements supplements nutrient, and supplement per week is primary later.After transplanting 23 days, 2MS moderate-element and micro member are supplemented
Element, supplement is primary within later 10 days.Transplanting 2 after half a month, transplants rooting rate up to 60% or more.
Embodiment 3: a kind of santal tissue-culturing rapid propagation micro cuttage method for culturing seedlings includes the following steps:
Exploiting entity: this laboratory cultures material is taken a little: the brilliant village Xian Yao
The culture of S1, rudiment bar: in October, 2015 selects 5 plants of select trees from the 8 years non-hibernating eggs source woodss in the brilliant village Xian Yao, selection
Illumination condition is good, the branch lopping immediate from the ground of robust growth, retains 40~60cm length, and the original in peace and quiet lopping
Coppice shoot.
S2, explant disinfection: in July, 2016 takes the select tree coppice shoot of 20~40cm, wipes out blade and stays petiole, uses in 12h
Ultrasonic drilling machine cleans 10min, and on super quiet workbench, sterilizes 13~20min with the mercury of 0.1mg/L, disinfection success rate reaches
60% or more.
S3, clump bud induction: the explant after disinfection accesses in clump bud inducement cultivation base, and Medium's PH Value is 5.8~6.2,
The element of MS including improvement 1, content are as follows: NH4NO3350~550mg/L, KH2PO4170~255mg/L, MgSO47H2O
370~550mg/L, KNO3 1900mg/L、CaCl2 400mg/L、Ca(NO3)27H2O 750mg/L, (NH4)2SO4260mg/L,
Microelement 1.5MS, organic matter 1.0MS, 0.10~0.2mg/L of hormone 6BA, IBA 0.2mg/L, 30~35mg/L of sucrose；
Squamous subculture condition be intensity of illumination be 2000~3000LX, 10 hour/day of light application time, cultivation temperature 22~28
℃。
S4, squamous subculture: after clump bud derives, it is transferred to santal subculture multiplication medium, condition of culture removes hormone concentration
Change it is outer, other conditions with step S3, expand it is numerous based on the squamous subculture period be 35 days, produced while expanding numerous seedling after being commissioned to train
Supporting the period is 45 days, and the hormone of subculture medium changes are as follows: hormone 6BA0.05~0.15mg/L, IBA0.10~0.15mg/L；
The same S3 of condition of culture.
S5, pre- culture of rootage: choosing height in squamous subculture, in 2.5cm or more, the sprout access that blade is sufficiently spread out is pre-
Root media carries out pre- culture of rootage and is transferred in experienced seedling shed after culture 25~35 days and practices seedling, improves 2 MS element and takes root in advance
Culture medium, the element comprising following content: NH4NO3560~1320mg/L, KH2PO4256~340mg/L, MgSO4 7H2O
148mg/L、KNO3760~1520mg/L, CaCl2 130mg/L、Ca(NO3)2 7H2O 750mg/L, microelement 1.0MS, have
Machine matter 1.0MS, hormone GGR 0.4mg/L, IBA 0.2mg/L, NAA 0.2mg/L, 40~50mg/L of sucrose.
S6, hardening culture: after taking root sprout culture 25~35 days in advance, it is transferred to the refining that intensity of illumination is 5000~10000Lx
Hardening in seedling greenhouse, the hardening time is 15~30 days, as sprout height >=4cm or more, transplants in time, reduces hardening time mistake
Long, pollution rate increases, and reduces bottle seedling bottle outlet rate；
S7, micro cuttage transplanting: use vermiculite: peat soil: yellow soil volume ratio is the micro cuttage matrix of 8:1:1, micro cuttage
It is preceding to sterilize matrix with 0.5% potassium permanganate.By taking root in advance and 0~5 day full lid film after the transplanting of the sprout of hardening, and cover two
The sunshade net of layer 75%, it is daily to open 4 times by spraying；6th~10 day opening film reservation sunshade net, 6 times/day of auto spraying, often
Secondary 10 seconds；The sunshade net of one layer 75% of reservation in 11st~20 day, 5 times/day of auto spraying, every time 10 seconds；Auto spraying 4 after 20 days
Times/day, 10 seconds every time.It is per week to carry out disease control using 1/1000 different fungicide.After transplanting 20 days, using 1/4MS
A great number of elements supplements nutrient, and supplement per week is primary later.After transplanting 23 days, 2MS moderate-element and microelement are supplemented, after
Supplement is primary within 10 days.Transplanting 2 after half a month, transplants rooting rate up to 60% or more.
Embodiment of above is only used to illustrate the technical scheme of the present invention and not to limit it, although referring to embodiment of above pair
The present invention is described in detail, those skilled in the art should understand that, technical solution of the present invention can be carried out
Modification or equivalent replacement should not all be detached from the spirit and scope of technical solution of the present invention.
Claims (5)
1. a kind of santal tissue-culturing rapid propagation micro cuttage method for culturing seedlings, which comprises the steps of:
The culture of S1, rudiment bar: in autumn and winter, according to tree height, the diameter of a cross-section of a tree trunk 1.3 meters above the ground in the fine provenance woods that the age of tree was more than 1/3 period of felling in turn
Choose select tree with the indexs such as symbiosis tree quantity, select illumination condition on select tree good, the branch closest to ground of robust growth into
Row lopping cutting handle, lopping retain 40~60cm length, remove the former coppice shoot in lopping, spring next year, germination rate was up to 95%；
S2, explant disinfection: in spring and summer, the 20~40cm long select tree coppice shoot sprouted after acquisition S1 processing is wiped out blade and is stayed
Petiole, 1~for 24 hours in 10min cleaned using ultrasonic drilling machine clear water, then on super quiet workbench, with the mercury disinfection 13 of 0.1g/L~
20min sterilizes success rate up to 60%；
S3, clump bud induction: the explant after disinfection accesses induction clump bud in clump bud inducement cultivation base and sprouts, and Medium's PH Value is
5.8~6.2, the element of the MS including improvement, content are as follows: NH4NO3350~550mg/L, KH2PO4170~255mg/L,
MgSO4.7H2O370~550mg/L, KNO3 1900mg/L、CaCl2 400mg/L、Ca(NO3)2.4H2O750mg/L、(NH4)2SO4260mg/L, microelement 1.5MS, organic matter 1.0MS, 0.10~0.2mg/L of hormone 6BA, IBA 0.2mg/L, sucrose
30~35mg/L；
S4, squamous subculture: after clump bud derives, it is transferred to santal subculture multiplication medium, condition of culture is changed except hormone concentration
Outside, other conditions are with step S3, breeding rate up to 2.5 times or more, expand it is numerous based on the squamous subculture period be 30~35 days, one
While expanding numerous squamous subculture period for producing seedling on one side is 40~50 days.The hormone of subculture medium changes are as follows: and hormone 6BA 0.05~
0.10~0.15mg/L of 0.15mg/L, IBA；
S5, pre- culture of rootage: choosing height in squamous subculture, in 2.5cm or more, the sprout access that blade is sufficiently spread out is taken root in advance
Culture medium carries out pre- culture of rootage and is transferred to hardening in hardening greenhouse after culture 25~35 days；
Pre- root media is the MS culture medium for improveing 2, content are as follows: NH4NO3560~1320mg/L, KH2PO4256~
340mg/L、MgSO4 7H2O 148mg/L、KNO3760~1520mg/L, CaCl2 130mg/L、Ca(NO3)2 4H2O
750mg/L, microelement 1.0MS, organic matter 1.0MS, hormone GGR 0.4mg/L, IBA 0.2mg/L, NAA 0.2mg/L, sugarcane
40~50mg/L of sugar.
S6, hardening culture: the pre- culture of rootage time is different because of season, is 25~35 days, then, without culture medium of transferring, directly
It is transferred to hardening in the hardening greenhouse that intensity of illumination is 5000~10000Lx, the hardening time is 15~30 days.When sprout height >=
It when 4cm or more, transplants in time, reduces hardening overlong time, pollution rate increases, and reduces bottle seedling bottle outlet rate；
S7, micro cuttage transplanting: it is suitable for the matrix formulations of pre- sprout micro cuttage transplanting of taking root: vermiculite: peat soil: the body of yellow soil
Product is than being 8:1:1.Matrix is sterilized with 0.5% potassium permanganate before micro cuttage, the different fungicide of use per week carry out after micro cuttage
Disease control.Because of the characteristics of santal Tube plantlets micro cuttage rootage duration is longer, and nutrition cannot be replenished in time in conventional fertilizer application, pre- life
After the sprout of root is transplanted 20 days, a great number of elements solution of 1 1/4MS~1/3MS of foliage-spray per week supplements nutrient.Transplanting 23
After it, every 10 days foliage-sprays, 1 1.5~2.0MS trace element solution.After the transplanting of santal Tube plantlets micro cuttage, early period is answered
Pay attention to moisturizing and how spraying, and cover sunshade net to reduce intensity of illumination, then gradually decrease spray timings and reinforces illumination.Micro- skewer
After inserting transplanting 2~2.5 months, rooting rate is transplanted up to 60% or more.
2. santal tissue-culturing rapid propagation micro cuttage method for culturing seedlings according to claim 1, which is characterized in that in step S1, setting
Age is more than that according to tree, the indexs such as the high, diameter of a cross-section of a tree trunk 1.3 meters above the ground and symbiosis tree quantity choose select tree in the fine provenance woods in 1/3 period of felling in turn, chooses select tree
The branch closest to ground of robust growth carries out lopping cutting handle, and lopping retains 40~60cm length, removes in lopping
Former coppice shoot, spring next year, germination rate was up to 95%.
3. santal tissue-culturing rapid propagation micro cuttage method for culturing seedlings according to claim 1, which is characterized in that acquisition is cut in step S2
The select tree coppice shoot of the 20~40cm sprouted after branch, 1~interior for 24 hours using ultrasonic drilling machine clear water cleaning 10min, then in super quiet workbench
On, 13~20min is sterilized with the mercury of 0.1g/L, sterilizes success rate up to 60% or more.
4. santal tissue-culturing rapid propagation micro cuttage method for culturing seedlings according to claim 1, which is characterized in that step S3, in S4, S5
Improvement 1, the minimal medium of improvement 2, hormone, the variation of sugar are unique, pass through the subculture that 1 culture medium of improvement obtains high breeding
Seedling obtains preculture of taking root by improvement 2 and does not fall leaves dead healthy and strong sprout.
5. santal tissue-culturing rapid propagation micro cuttage method for culturing seedlings according to claim 1, which is characterized in that step S6, needle in S7
And take root difficult, sprout slow to santal growth is easy dehydration and is easy again by the spy of germ dip dyeing, incubation time length, nutritional supplementation hardly possible
Point, being equipped with the grafting matrix that optimal reduction germ is infected is micro cuttage grafting matrix (volume ratio are as follows: vermiculite: peat soil: gold zone
Soil=8:1:1), in conjunction with specific a great number of elements and microelement foliar spray complementary technology measure, the effective moisture applied with
The control of germ series realizes that micro cuttage transplants rooting rate up to 60% or more.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710718937.6A CN109744142A (en) | 2017-08-21 | 2017-08-21 | A kind of santal tissue-culturing quick-propagation method for culturing seedlings |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710718937.6A CN109744142A (en) | 2017-08-21 | 2017-08-21 | A kind of santal tissue-culturing quick-propagation method for culturing seedlings |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109744142A true CN109744142A (en) | 2019-05-14 |
Family
ID=66397611
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710718937.6A Pending CN109744142A (en) | 2017-08-21 | 2017-08-21 | A kind of santal tissue-culturing quick-propagation method for culturing seedlings |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109744142A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110122084A (en) * | 2019-05-23 | 2019-08-16 | 宁德师范学院 | Cuttage breeding method in a kind of Morinda officinalis tissue-cultured seedling room |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH08298887A (en) * | 1995-05-08 | 1996-11-19 | Yamashita Yukinori | Tissue culture of santalum and its secondary proliferation |
| CN101213936A (en) * | 2007-12-29 | 2008-07-09 | 中国科学院华南植物园 | Tissue culture fast breeding method for sandalwood |
| CN101983557A (en) * | 2010-10-19 | 2011-03-09 | 廖俊杰 | In vitro quick breeding method of seedling stem of santal seed embryo |
| CN105123523A (en) * | 2015-09-07 | 2015-12-09 | 中国林业科学研究院热带林业研究所 | Albizia falcataria forest tree selecting fast seedling growing method |
-
2017
- 2017-08-21 CN CN201710718937.6A patent/CN109744142A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH08298887A (en) * | 1995-05-08 | 1996-11-19 | Yamashita Yukinori | Tissue culture of santalum and its secondary proliferation |
| CN101213936A (en) * | 2007-12-29 | 2008-07-09 | 中国科学院华南植物园 | Tissue culture fast breeding method for sandalwood |
| CN101983557A (en) * | 2010-10-19 | 2011-03-09 | 廖俊杰 | In vitro quick breeding method of seedling stem of santal seed embryo |
| CN105123523A (en) * | 2015-09-07 | 2015-12-09 | 中国林业科学研究院热带林业研究所 | Albizia falcataria forest tree selecting fast seedling growing method |
Non-Patent Citations (4)
| Title |
|---|
| B JANARTHANAUM等: "High Frequency Shoot Regeneration from Internodal Explants of Santalum album L.", 《INTERNATIONAL JOURNAL OF BOTANY 》 * |
| C. K. SINGH等: "Plant regeneration from leaf explants of mature sandalwood (Santalum album L.) trees under in vitro conditions", 《IN VITRO CELL.DEV.BIOL.—PLANT》 * |
| C.K.SINGH等: "Effect of plant growth regulators on in vitro plant regeneration of sandalwood (Santalum album L.) via organogenesis", 《AGROFOREST SYST》 * |
| 马国华等: "檀香的组织培养与快速繁殖 ", 《植物生理学通讯》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110122084A (en) * | 2019-05-23 | 2019-08-16 | 宁德师范学院 | Cuttage breeding method in a kind of Morinda officinalis tissue-cultured seedling room |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102499090B (en) | Method for isolated culture of Haworthia succulent plants | |
| CN103988777B (en) | A kind of in-vitro culture method for tender stem segments of the wide yulan of leaflet dwarf form | |
| CN101946709B (en) | Seedling raising method of plateau rhodiola crenulata | |
| CN104885773A (en) | Method for rapidly cultivating early shaping tissue culture commodity seedlings of blueberries | |
| CN104273037B (en) | A kind of tissue culture and rapid propagation method of tail alpine ash DH33-27 kind | |
| CN104285814B (en) | A kind of tissue culture and rapid propagation method of tail alpine ash DH32-43 kind | |
| CN104273030B (en) | Caulis Sacchari sinensis test tube Seedling photoautotrophy rooting method | |
| CN103202232A (en) | Method for efficiently and industrially producing sweet potato detoxification tissue culture seedlings | |
| CN104472366A (en) | Tissue culture rapid-propagation method for improving salt tolerance of seedlings of southern ecotype jujubes | |
| CN101828508B (en) | Method for rapidly promoting assimilation of nitrate in vegetables planted by soilless culture technology | |
| CN107018896A (en) | A kind of method of facility cuttage tilia miqueliana | |
| CN105359948B (en) | Hybrid larch micro cuttage method for culturing seedlings | |
| CN103636374A (en) | Light supplement seedling culture method for Lijiang spruce container seedlings and application thereof | |
| CN102499034B (en) | Soilless cultivation method capable of reducing content of nitrate in curled mallow and curled mallow nutrient solution | |
| CN101563982A (en) | Camellia seedling method for quick growth | |
| CN103843650B (en) | Casuarina equisetifolia L. water planting method for culturing seedlings | |
| CN106376461B (en) | The method of China fir tissue-cultured seedling culture of rootage and hardening | |
| CN102613089B (en) | High-efficient in-vitro propagation method of 20-year-old schima superba big tree | |
| CN104285815A (en) | Tissue culture and rapid propagation method of E. urophylla*E. Grandis DH32-13 | |
| CN104304028B (en) | A kind of tissue culture and rapid propagation method of tail alpine ash DH32-26 kind | |
| CN111492978A (en) | Tissue culture rapid propagation method of Eucalyptus macrocarpa 1212 variety | |
| CN103477989A (en) | Method for cultivating eucalyptus tissue culture seedling by adopting light media container | |
| CN105145042B (en) | Virus-free basic potato seed soil incubation method | |
| CN103869058A (en) | Rapid evaluation method for salt tolerance of tomatoes | |
| CN111492974A (en) | Tissue culture rapid propagation method of Eucalyptus robusta 1204 variety |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |