CN103718963B - A kind of cultural method of horseradish detoxic seedling - Google Patents
A kind of cultural method of horseradish detoxic seedling Download PDFInfo
- Publication number
- CN103718963B CN103718963B CN201310724001.6A CN201310724001A CN103718963B CN 103718963 B CN103718963 B CN 103718963B CN 201310724001 A CN201310724001 A CN 201310724001A CN 103718963 B CN103718963 B CN 103718963B
- Authority
- CN
- China
- Prior art keywords
- horseradish
- illumination
- hours
- condition
- sprout
- 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.)
- Expired - Fee Related
Links
- 240000003291 Armoracia rusticana Species 0.000 title claims abstract description 50
- 235000011330 Armoracia rusticana Nutrition 0.000 title claims abstract description 44
- 238000000034 method Methods 0.000 title claims abstract description 18
- 238000010792 warming Methods 0.000 claims abstract description 15
- 230000004069 differentiation Effects 0.000 claims abstract description 9
- 230000000442 meristematic effect Effects 0.000 claims abstract description 8
- 238000005286 illumination Methods 0.000 claims description 52
- 230000003203 everyday effect Effects 0.000 claims description 26
- 239000002689 soil Substances 0.000 claims description 16
- 229910052799 carbon Inorganic materials 0.000 claims description 11
- 239000008223 sterile water Substances 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 241000196324 Embryophyta Species 0.000 claims description 9
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 7
- 238000004659 sterilization and disinfection Methods 0.000 claims description 7
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 claims description 6
- 235000000760 Wasabia japonica Nutrition 0.000 claims description 6
- 244000195452 Wasabia japonica Species 0.000 claims description 6
- 230000001954 sterilising effect Effects 0.000 claims description 6
- TWFZGCMQGLPBSX-UHFFFAOYSA-N Carbendazim Natural products C1=CC=C2NC(NC(=O)OC)=NC2=C1 TWFZGCMQGLPBSX-UHFFFAOYSA-N 0.000 claims description 5
- JNPZQRQPIHJYNM-UHFFFAOYSA-N carbendazim Chemical compound C1=C[CH]C2=NC(NC(=O)OC)=NC2=C1 JNPZQRQPIHJYNM-UHFFFAOYSA-N 0.000 claims description 5
- 239000006013 carbendazim Substances 0.000 claims description 5
- 229960002523 mercuric chloride Drugs 0.000 claims description 5
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 claims description 5
- 235000016709 nutrition Nutrition 0.000 claims description 5
- 230000035764 nutrition Effects 0.000 claims description 5
- 239000012286 potassium permanganate Substances 0.000 claims description 5
- 230000001737 promoting effect Effects 0.000 claims description 5
- 229910052902 vermiculite Inorganic materials 0.000 claims description 5
- 235000019354 vermiculite Nutrition 0.000 claims description 5
- 239000010455 vermiculite Substances 0.000 claims description 5
- 230000006698 induction Effects 0.000 claims description 2
- 239000000463 material Substances 0.000 abstract description 4
- 238000009395 breeding Methods 0.000 abstract description 2
- 230000001488 breeding effect Effects 0.000 abstract description 2
- 239000000835 fiber Substances 0.000 abstract description 2
- 230000004083 survival effect Effects 0.000 description 9
- 201000010099 disease Diseases 0.000 description 8
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 8
- 241000700605 Viruses Species 0.000 description 7
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 5
- 238000003745 diagnosis Methods 0.000 description 5
- 230000008030 elimination Effects 0.000 description 5
- 238000003379 elimination reaction Methods 0.000 description 5
- 238000003118 sandwich ELISA Methods 0.000 description 5
- 230000003612 virological effect Effects 0.000 description 5
- 229920001817 Agar Polymers 0.000 description 4
- 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 4
- 229930006000 Sucrose Natural products 0.000 description 4
- 239000008272 agar Substances 0.000 description 4
- 239000005720 sucrose Substances 0.000 description 4
- 238000012258 culturing Methods 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 210000003462 vein Anatomy 0.000 description 3
- 240000003945 Cochlearia officinalis Species 0.000 description 2
- 241000918585 Pythium aphanidermatum Species 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 235000007463 Cochlearia officinalis Nutrition 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 241000243785 Meloidogyne javanica Species 0.000 description 1
- 241000233679 Peronosporaceae Species 0.000 description 1
- 208000004880 Polyuria Diseases 0.000 description 1
- 244000184734 Pyrus japonica Species 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000002924 anti-infective effect Effects 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000035619 diuresis Effects 0.000 description 1
- 235000018927 edible plant Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- DNJIEGIFACGWOD-UHFFFAOYSA-N ethanethiol Chemical compound CCS DNJIEGIFACGWOD-UHFFFAOYSA-N 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 208000006278 hypochromic anemia Diseases 0.000 description 1
- -1 isosulfocyanate compound Chemical class 0.000 description 1
- 235000019508 mustard seed Nutrition 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 230000007226 seed germination Effects 0.000 description 1
- 108010058651 thioglucosidase Proteins 0.000 description 1
Landscapes
- Cultivation Of Plants (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a kind of cultural method of horseradish detoxic seedling, the method selects aseptic horseradish sprout to be culture materials, after intermittent warming, get in its bud meristematic tissue access differentiation medium, again through strong seedling culture base and root media Fiber differentiation and obtain horseradish detoxic seedling, the horseradish detoxic seedling cultivated has fast growth, active principle content advantages of higher.The inventive method is that the quickly breeding of horseradish improved seeds provides a new way.
Description
Technical field
The present invention relates to the method for tissue culture of a kind of horseradish plant, particularly relate to a kind of cultural method of horseradish detoxic seedling.
Background technology
Horseradish (Wasabi japonica Matsum) has another name called scurvy grass, mountain dish, is Cruciferae scurvy grass, belongs to perennial half cloudy raw herbaceous plant.Material containing a kind of mustard seed glycoside in horseradish, when plant corpus is grated, this material is hydrolyzed and forms isosulfocyanate compound under the effect of myrosinase, there is sterilization, anticorrosion, diuresis, purify the blood, the multiple medical active such as anti-infective, anticancer, be the very rare and precious medicinal and edible plant of generally acknowledging in the world.
Horseradish plant grows the harm being often subject to many damage by disease and insect under field conditions (factors), ink is had to enter the multiple Common Diseases symptoms such as disease, downy mildew, white blister, damping off, soft rot, virus disease and root knot nematode disease as reported horseradish plant specially in the Research Literature " Yunnan Province's horseradish Common Diseases and symptoms " of the people such as He Yueqiu, wherein enter with ink the harm that disease causes the leaf of horseradish, petiole, rhizome, coring maximum, directly cause Quality Down and the underproduction of horseradish product, horseradish plant time serious, also can be caused all dead; Damping off main harm seedling base portion, is not yet unearthed dead with regard to browning sometimes after seed germination; The ill plant of virus disease shows as chlorosis between blade most vein, the floral leaf shape alternate in yellow green, and some blades diminish, deformity, and vein has bright arteries and veins phenomenon to light, and plant growing way is bad; Soft rot main harm rhizome and petiole, at the rhizome harm initial stage, peripheral blade shows wilting under burning sun, rhizome interior tissue disintegration time serious, produces the bacterial ooze of canescence stench, and last whole strain is wilted dead.Therefore, for keeping the fine quality of horseradish plant, and improving its output, being necessary the method for tissue culture that a kind of horseradish plant is provided.
Summary of the invention
The object of the present invention is to provide a kind of cultural method of horseradish detoxic seedling.
The cultural method of horseradish detoxic seedling provided by the invention comprises the steps:
(1) cut the bud nose part of the sprout that Wasabi roots grows, place it on superclean bench, first sterilize 2 ~ 5 minutes with the mercuric chloride of 0.1%, then with after sterile water wash, be inoculated in the medium MS+KT0.5 ~ 1.5mgL promoting that bud point grows fast
-1+ NAA0.3 ~ 1.0mgL
-1in, 12 ~ 20 DEG C, illumination every day 6 ~ 12 hours, intensity of illumination cultivate under being the condition of 1000 ~ 1500lx;
(2) length of cultivating in selecting step (1), the aseptic sprout of more than 1.0cm, is transferred into MS+6-BA1 ~ 3.0mgL
-1+ IAA0.5 ~ 2.0mgL
-1in the medium of+proline 0.5 ~ 2%+ active carbon 1 ~ 3%, first cultivate 15 days under the condition of 5 DEG C, then be placed in incubator the intermittent warming carried out 8 ~ 15 days;
(3) choose well-grown sprout under anatomical lens, after carefully peelling off the spire outside sprout with dissecting needle, cut the bud meristematic tissue with 1 ~ 4 leaf primordium, access differentiation medium MS+6-BA1.0 ~ 3mgL
-1+ KT0.1 ~ 0.5mgL
-1in, 15 ~ 22 DEG C, illumination every day 10 ~ 14 hours, intensity of illumination cultivate under being the condition of 1000 ~ 1500lx;
(4) choose the horseradish Multiple Buds of growth, after being divided into individual plant, proceed to strong seedling culture base MS+NAA0.05 ~ 0.1mgL
-1+ 6-BA0.1 ~ 0.5mgL
-1+ KT0.05 ~ 0.2mgL
-1in, 10 ~ 18 DEG C, illumination every day 10 ~ 16 hours, intensity of illumination cultivate under being the condition of 1000 ~ 1500lx;
(5) horseradish of robust growth is chosen without offspring access root media White+NAA0.1 ~ 1.0mgL
-1in, 15 ~ 22 DEG C, illumination every day 6 ~ 10 hours, intensity of illumination be the condition of 800 ~ 1000lx under root induction;
(6) when growing more than 3 ~ 4 adventive root without offspring, open bottle cap, after 2 ~ 4 days, it is taken out from blake bottle in indoor hardening, wash away the medium on adventive root with sterile water after, transplant in the soil to sterilization and grow.
Further, the bud nose part of sprout described in step (1) refers to that sprout upper length is the bud nose part of 0.2 ~ 0.5cm.
Further, intermittent warming described in step (2) refer to cultivate under the condition of 8 ~ 12 DEG C 19 hours with under the condition of 30 ~ 45 DEG C, cultivate 5 hours hocket.
Further, soil described in step (6) be by vermiculite and full Nutrition Soil formulated by 2:1, soil disinfection refer to first adopt 0.1% carbendazim sterilization 6 ~ 10 hours, then with 0.1% potassium permanganate sterilize 12 ~ 24 hours.
Further, the condition of culture of hardening is temperature 15 ~ 20 DEG C and relative moisture 80% ~ 95% in step (6), and the condition of culture after transplanting is cultivation temperature 12 ~ 20 DEG C, relative moisture is 70 ~ 80%, illumination every day 4 ~ 10 hours and intensity of illumination are 800 ~ 1200lx.
The pH value of above-mentioned all medium is 5.6 ~ 6.2, agar 5.5 ~ 6.5gL
-1, sucrose 25 ~ 35gL
-1.
The present invention selects aseptic horseradish sprout to be culture materials, after intermittent warming, get in its bud meristematic tissue access differentiation medium, again through strong seedling culture base and root media Fiber differentiation and obtain horseradish detoxic seedling, the horseradish detoxic seedling cultivated has fast growth, active principle content advantages of higher.The inventive method is that the quickly breeding of horseradish improved seeds provides a new way.
Below in conjunction with embodiment, the present invention is described in further detail.
Embodiment
Embodiment 1
(1) the bud nose part that the sprout upper length that Wasabi roots grows is 0.2cm is cut, place it on superclean bench, first to sterilize 2 minutes with the mercuric chloride of 0.1%, then after using sterile water wash 4 times, be inoculated in the medium MS+KT0.5mgL promoting the growth fast of bud point
-1+ NAA0.3mgL
-1in, cultivate 25 days under temperature 12 DEG C, illumination every day 6 hours, intensity of illumination are the condition of 1000lx, the aseptic sprout growing to more than 1.0cm has 89.32%;
(2) length of cultivating in selecting step (1), the aseptic sprout of more than 1.0cm, is transferred into MS+6-BA1.0mgL
-1+ IAA0.5mgL
-1in the medium of+proline 0.5%+ active carbon 1.0%, first be placed in the refrigerator Low-temperature culture 15 days of 5 DEG C, be placed in incubator the intermittent warming carried out 8 days again, described intermittent warming refer to cultivate under the condition of 8 DEG C 19 hours with under the condition of 30 DEG C, cultivate 5 hours hocket, the survival rate of sprout is 98.68%;
(3) choose well-grown sprout after step (2) process and, under anatomical lens, after carefully peelling off the spire outside sprout with dissecting needle, cut the bud meristematic tissue with 1 leaf primordium, access differentiation medium MS+6-BA1.0mgL
-1+ KT0.1mgL
-1in, cultivate 30 days under temperature 15 DEG C, illumination every day 14 hours, intensity of illumination are the condition of 1000lx, the inductivity of Multiple Buds is 32.53%, and adopt double-antibodies sandwich ELISA to carry out Viral diagnosis the horseradish Multiple Buds of cultivation, its virus elimination rate is 96.46%;
(4) choose the horseradish Multiple Buds of growth, after being divided into individual plant, proceed to strong seedling culture base MS+NAA0.05mgL
-1+ 6-BA0.1mgL
-1+ KT0.05mgL
-1in, cultivate 25 days under temperature 10 DEG C, illumination every day 10 hours, intensity of illumination are the condition of 1000lx;
(5) horseradish of the robust growth of cultivating in selecting step (4) is chosen without offspring access root media White+NAA0.1mgL
-1in, cultivate 30 days under temperature 15 DEG C, illumination every day 6 hours, intensity of illumination are the condition of 800lx, horseradish is 84.63% without the rooting rate of offspring;
(6) when growing more than 3 adventive root without offspring, open bottle cap, first temperature to be 15 DEG C and relative moisture be 80% culturing room in hardening 2 days, again it is taken out from blake bottle, wash away the medium on adventive root with sterile water after, transplant to growing by the soil that 0.1% carbendazim is sterilized 6 hours and 0.1% potassium permanganate is sterilized 12 hours, described soil be by vermiculite and full Nutrition Soil formulated by 2:1; Detoxic seedling grows under temperature 12 DEG C, relative moisture 70%, illumination every day 4 hours and intensity of illumination are the condition of 800lx, and the survival rate of detoxic seedling is 92.62%;
The pH value of above-mentioned all medium is 5.6, agar 5.5gL
-1, sucrose 25gL
-1.
Embodiment 2
(1) the bud nose part that the sprout upper length that Wasabi roots grows is 0.3cm is cut, place it on superclean bench, first to sterilize 3 minutes with the mercuric chloride of 0.1%, then after using sterile water wash 5 times, be inoculated in the medium MS+KT1.0mgL promoting the growth fast of bud point
-1+ NAA0.5mgL
-1in, carry out cultivation 25 days in temperature 18 DEG C, illumination every day 8 hours, intensity of illumination under being the condition of 1200lx, the aseptic sprout growing to more than 1.0cm has 92.26%;
(2) length of cultivating in selecting step (1), the aseptic sprout of more than 1.0cm, is transferred into MS+6-BA2.0mgL
-1+ IAA1.5mgL
-1in the medium of+proline 1.2%+ active carbon 2.0%, first be placed in the refrigerator Low-temperature culture 15 days of 5 DEG C, be placed in incubator the intermittent warming carried out 12 days again, described intermittent warming refers to cultivate at 10 DEG C and within 5 hours, hockets with cultivating at 35 DEG C for 19 hours, and the survival rate of sprout is 96.57%;
(3) choose well-grown sprout after step (2) process and, under anatomical lens, after carefully peelling off the spire outside sprout with dissecting needle, cut the bud meristematic tissue with 2 leaf primordium, access differentiation medium MS+6-BA2.0mgL
-1+ KT0.3mgL
-1in, cultivate 30 days under temperature 18 DEG C, illumination every day 12 hours, intensity of illumination are the condition of 1200lx, the inductivity of Multiple Buds is 51.42%; Adopt double-antibodies sandwich ELISA to carry out Viral diagnosis the horseradish Multiple Buds of cultivation, its virus elimination rate is 94.76%;
(4) choose the horseradish Multiple Buds of growth, after being divided into individual plant, proceed to strong seedling culture base MS+NAA0.08mgL
-1+ 6-BA0.3mgL
-1+ KT0.1mgL
-1in, cultivate 25 days under temperature 15 DEG C, illumination every day 12 hours, intensity of illumination are the condition of 1200lx;
(5) horseradish of the robust growth of cultivating in selecting step (4) is without offspring access root media White+NAA0.5mgL
-1in, cultivate 30 days under temperature 18 DEG C, illumination every day 8 hours, intensity of illumination are the condition of 900lx, horseradish is 92.13% without the rooting rate of offspring;
(6) when growing more than 4 adventive root without offspring, open bottle cap, first temperature to be 18 DEG C and relative moisture be 88% culturing room in hardening 3 days, again it is taken out from blake bottle, wash away the medium on adventive root with sterile water after, transplant to growing by the soil that 0.1% carbendazim is sterilized 8 hours and 0.1% potassium permanganate is sterilized 18 hours, described soil be by vermiculite and full Nutrition Soil formulated by 2:1; Detoxic seedling grows under temperature 16 DEG C, relative moisture 75%, illumination every day 6 hours and intensity of illumination are the condition of 1000lx, and the survival rate of detoxic seedling is 98.37%;
The pH value of above-mentioned all medium is 5.8, agar 6.0gL
-1, sucrose 30gL
-1.
Embodiment 3
(1) the bud nose part that the sprout upper length that Wasabi roots grows is 0.5cm is cut, place it on superclean bench, first to sterilize 5 minutes with the mercuric chloride of 0.1%, then after using sterile water wash 6 times, be inoculated in the medium MS+KT1.5mgL promoting the growth fast of bud point
-1+ NAA1.0mgL
-1in, carry out cultivation 25 days in temperature 20 DEG C, illumination every day 12 hours, intensity of illumination under being the condition of 1500lx, the aseptic sprout growing to more than 1.0cm has 100%;
(2) length of cultivating in selecting step (1), the aseptic sprout of more than 1.0cm, is transferred into MS+6-BA3.0mgL
-1+ IAA2.0mgL
-1in the medium of+proline 2%+ active carbon 3%, first be placed in the refrigerator Low-temperature culture 15 days of 5 DEG C, be placed in incubator the intermittent warming carried out 15 days again, described intermittent warming refer to cultivate under the condition of 12 DEG C 19 hours with under the condition of 45 DEG C, cultivate 5 hours hocket, the survival rate of sprout is 83.36%;
(3) choose well-grown sprout after step (2) process and, under anatomical lens, after carefully peelling off the spire outside sprout with dissecting needle, cut the bud meristematic tissue with 4 leaf primordium, access differentiation medium MS+6-BA3.0mgL
-1+ KT0.5mgL
-1in, cultivate 30 days under temperature 22 DEG C, illumination every day 14 hours, intensity of illumination are the condition of 1500lx, the inductivity of Multiple Buds is 76.45%; Adopt double-antibodies sandwich ELISA to carry out Viral diagnosis the horseradish Multiple Buds of cultivation, its virus elimination rate is 73.52%;
(4) choose the horseradish Multiple Buds of growth, after being divided into individual plant, proceed to strong seedling culture base MS+NAA0.1mgL
-1+ 6-BA0.5mgL
-1+ KT0.2mgL
-1in, cultivate 25 days under temperature 18 DEG C, illumination every day 16 hours, intensity of illumination are the condition of 1500lx;
(5) horseradish of the robust growth of cultivating in selecting step (4) is without offspring access root media White+NAA1.0mgL
-1in, cultivate 30 days under temperature 22 DEG C, illumination every day 10 hours, intensity of illumination are the condition of 1000lx, horseradish is 89.21% without the rooting rate of offspring;
(6) when growing more than 4 adventive root without offspring, open bottle cap, first temperature to be 20 DEG C and relative moisture be 95% culturing room in hardening 4 days, again it is taken out from blake bottle, wash away the medium on adventive root with sterile water after, transplant to growing by the soil that 0.1% carbendazim is sterilized 10 hours and 0.1% potassium permanganate is sterilized 24 hours, described soil be by vermiculite and full Nutrition Soil formulated by 2:1; Detoxic seedling grows under temperature 20 DEG C, relative moisture 80%, illumination every day 10 hours and intensity of illumination are the condition of 1200lx, and the survival rate of detoxic seedling is 97.16%;
The pH value of above-mentioned all medium is 6.2, agar 6.5gL
-1, sucrose 35gL
-1.
Embodiment 4
To promote described in embodiment 2 step (1) that the medium that bud point grows fast changes MS+KT0.5mgL into
-1+ NAA0.8mgL
-1, other step is with embodiment 2, and cultivate after 25 days, the aseptic sprout growing to more than 1.0cm has 90.57%.
Embodiment 5
Change medium described in embodiment 2 step (2) into MS+6-BA1.0mgL
-1+ IAA1.5mgL
-1+ proline 1.2%+ active carbon 2.0%, the time of intermittent warming changes 10 days into, and other steps are with embodiment 2, and the survival rate of sprout is 94.53%.
Embodiment 6
The intermittent warming mode of the aseptic sprout of horseradish in embodiment 2 step (2) is changed into and under the condition of 8 DEG C, to cultivate 19h and under 40 DEG C of conditions, cultivate 5h hocket, other steps are with embodiment 2, the horseradish Multiple Buds double-antibodies sandwich ELISA of cultivating carries out Viral diagnosis, and its virus elimination rate is 95.06%.
Embodiment 7
The bud meristematic tissue reconfiguration of leaf primordium described in embodiment 2 step (3) is entered MS+6-BA1mgL
-1+ KT0.5mgL
-1differentiation medium in, temperature 16 DEG C, illumination every day 13 hours and intensity of illumination are cultivate 30 days under the condition of 1000lx, and other steps are with embodiment 2, and the inductivity of Multiple Buds is 49.82%.
Comparing embodiment 1
Change medium described in embodiment 2 step (2) into MS+6-BA2.0mgL
-1+ 2.4-D2.0mgL
-1+ active carbon 2.0%, other steps are with embodiment 2, and the survival rate of sprout is 57.86%.
Comparing embodiment 2
The intermittent warming mode of the aseptic sprout of horseradish in embodiment 2 step (2) changed into and under the condition of 5 DEG C, to cultivate 5h and under 45 DEG C of conditions, cultivate 19h hocket, other steps are with embodiment 2, and the survival rate of sprout is 12.56%.
Comparing embodiment 3
The step (2) of embodiment 2 cancelled, other steps are with embodiment 2, and the horseradish Multiple Buds double-antibodies sandwich ELISA of cultivation carries out Viral diagnosis, and its virus elimination rate is 31.43%.
Comparing embodiment 4
The root media described in embodiment 2 is changed into MS+NAA0.5mgL by embodiment 2 step (5)
-1, other steps are with embodiment 2, and horseradish is 79.73% without the rooting rate of offspring.
Claims (4)
1. a cultural method for horseradish detoxic seedling, is characterized in that comprising the steps:
(1) cut the bud nose part of the sprout that Wasabi roots grows, place it on superclean bench, first sterilize 2 ~ 5 minutes with the mercuric chloride of 0.1%, then with after sterile water wash, be inoculated in the medium MS+KT0.5 ~ 1.5mgL promoting that bud point grows fast
-1+ NAA0.3 ~ 1.0mgL
-1in, 12 ~ 20 DEG C, illumination every day 6 ~ 12 hours, intensity of illumination cultivate under being the condition of 1000 ~ 1500lx;
(2) length of cultivating in selecting step (1), the aseptic sprout of more than 1.0cm, is transferred into MS+6-BA 1.0 ~ 3.0mgL
-1+ IAA0.5 ~ 2.0mgL
-1in the medium of+proline 0.5 ~ 2%+ active carbon 1 ~ 3%, first cultivate 15 days under the condition of 5 DEG C, be placed in incubator the intermittent warming carried out 8 ~ 15 days again, described intermittent warming refer to cultivate under the condition of 8 ~ 12 DEG C 19 hours with under the condition of 30 ~ 45 DEG C, cultivate 5 hours hocket;
(3) choose well-grown sprout under anatomical lens, after carefully peelling off the spire outside sprout with dissecting needle, cut the bud meristematic tissue with 1 ~ 4 leaf primordium, access differentiation medium MS+6-BA1 ~ 3mgL
-1+ KT0.1 ~ 0.5mgL
-1in, 15 ~ 22 DEG C, illumination every day 10 ~ 14 hours, intensity of illumination cultivate under being the condition of 1000 ~ 1500lx;
(4) choose the horseradish Multiple Buds of growth, after being divided into individual plant, proceed to strong seedling culture base MS+NAA0.05 ~ 0.1mgL
-1+ 6-BA0.1 ~ 0.5mgL
-1+ KT0.05 ~ 0.2mgL
-1in, 10 ~ 18 DEG C, illumination every day 10 ~ 16 hours, intensity of illumination cultivate under being the condition of 1000 ~ 1500lx;
(5) horseradish of robust growth is chosen without offspring access root media White+NAA0.1 ~ 1.0mgL
-1in, 15 ~ 22 DEG C, illumination every day 6 ~ 10 hours, intensity of illumination be the condition of 800 ~ 1000lx under root induction;
(6) when growing more than 3 ~ 4 adventive root without offspring, open bottle cap, after 2 ~ 4 days, it is taken out from blake bottle in indoor hardening, wash away the medium on adventive root with sterile water after, transplant in the soil to sterilization and grow.
2. the cultural method of a kind of horseradish detoxic seedling according to claim 1, is characterized in that: the bud nose part of sprout described in step (1) refers to that sprout upper length is the bud nose part of 0.2 ~ 0.5cm.
3. the cultural method of a kind of horseradish detoxic seedling according to claim 1, it is characterized in that: soil described in step (6) be by vermiculite and full Nutrition Soil formulated by 2:1, sterilization refers to and first adopts 0.1% carbendazim sterilization 6 ~ 10 hours, then sterilizes 12 ~ 24 hours with 0.1% potassium permanganate.
4. the cultural method of a kind of horseradish detoxic seedling according to claim 1, it is characterized in that: the condition of culture of hardening is temperature 15 ~ 20 DEG C and relative moisture 80% ~ 95% in step (6), the condition of culture after transplanting is cultivation temperature 12 ~ 20 DEG C, relative moisture is 70 ~ 80%, illumination every day 4 ~ 10 hours and intensity of illumination be 800 ~ 1200lx.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310724001.6A CN103718963B (en) | 2013-12-24 | 2013-12-24 | A kind of cultural method of horseradish detoxic seedling |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310724001.6A CN103718963B (en) | 2013-12-24 | 2013-12-24 | A kind of cultural method of horseradish detoxic seedling |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103718963A CN103718963A (en) | 2014-04-16 |
| CN103718963B true CN103718963B (en) | 2015-07-29 |
Family
ID=50443460
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310724001.6A Expired - Fee Related CN103718963B (en) | 2013-12-24 | 2013-12-24 | A kind of cultural method of horseradish detoxic seedling |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103718963B (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1204453A (en) * | 1998-08-05 | 1999-01-13 | 南京农业大学 | Method for quick reproducing 'Shankui' sprout |
| JP2957102B2 (en) * | 1994-12-02 | 1999-10-04 | ハウス食品株式会社 | Enlargement culture method of wasabi rhizome |
| CN102823491A (en) * | 2012-08-06 | 2012-12-19 | 成都大学 | Method for culturing horseradish callus through taking horseradish root as explant |
| CN103053426A (en) * | 2013-02-04 | 2013-04-24 | 四川蓝公府农业科技有限公司 | Horseradish plantlet culture method |
| CN103125386A (en) * | 2013-02-04 | 2013-06-05 | 四川蓝公府农业科技有限公司 | Industrial horseradish planting method |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6485016A (en) * | 1987-09-24 | 1989-03-30 | New Bio Giken Kk | Aquiculture of japanese horseradish |
| KR100674115B1 (en) * | 2005-04-29 | 2007-01-30 | 주식회사 비트로시스 | Method for mass-producing wasabia and its root |
-
2013
- 2013-12-24 CN CN201310724001.6A patent/CN103718963B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2957102B2 (en) * | 1994-12-02 | 1999-10-04 | ハウス食品株式会社 | Enlargement culture method of wasabi rhizome |
| CN1204453A (en) * | 1998-08-05 | 1999-01-13 | 南京农业大学 | Method for quick reproducing 'Shankui' sprout |
| CN102823491A (en) * | 2012-08-06 | 2012-12-19 | 成都大学 | Method for culturing horseradish callus through taking horseradish root as explant |
| CN103053426A (en) * | 2013-02-04 | 2013-04-24 | 四川蓝公府农业科技有限公司 | Horseradish plantlet culture method |
| CN103125386A (en) * | 2013-02-04 | 2013-06-05 | 四川蓝公府农业科技有限公司 | Industrial horseradish planting method |
Non-Patent Citations (6)
| Title |
|---|
| Biochemical markers of morphogenesis in long term horseradish (Armoracia lapathifolia Gilib.) tissue culture;Biljana Balen et al.;《Scientia Horticulturae》;20091231;第119卷;第88-97页 * |
| Plant Regeneration from Leaf Blade Explants of Horseradish (Amoracia rusticana L.) through In Vitro Culture;Erwin Yuliadi;《Jurnal Akta Agrosia》;20080630;第11卷(第1期);第63-68页 * |
| 不同激素种类和浓度对山葵(Eutrema wasabi)组培苗增殖效果的研究;吴震等;《西南农业学报》;20021231;第15卷(第3期);第66-69页 * |
| 山葵的离体培养和植株再生;王广东等;《植物生理学通讯 》;19980831;第34卷(第4期);第267页 * |
| 山葵的组织培养和快速繁殖;刘琴等;《植物生理学通讯》;20021231;第38卷(第6期);第581页 * |
| 日本山葵的组培快繁试验研究;林加根等;《现代农业科技》;20081231(第21期);第24-25页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103718963A (en) | 2014-04-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101611697B (en) | Virus removal and rapid propagation technology of sweet potato variety 'Shangshu 19' | |
| CN104106468B (en) | The quick breeding method for tissue culture of a kind of radix fici simplicissimae | |
| CN103931497B (en) | A kind of method improving dragon fruit plantlet in vitro planting percent | |
| CN104272981B (en) | A kind of grafting method of tomato | |
| CN103766222B (en) | Sweet Stevia tissue culture method and substratum thereof | |
| CN104303851A (en) | Tomato grafting method capable of improving survival rate | |
| CN102907323A (en) | Method for aseptically producing miniature seed stems of common bletilla pseudobulb seeds | |
| CN102845313A (en) | Method for quickly in-vitro actinidia kolomikta propagating | |
| CN101129126A (en) | Cottage root promoting method of blueberry green branch | |
| CN102217550A (en) | Fast-propagation technology and composition of culture medium for virus-free plantlets of red bud taros | |
| CN101897297B (en) | Two-step tissue culture quick propagation method for hemerocallis | |
| CN101926284B (en) | Monkshood-tuber tissue culture and rapid propagation method | |
| CN101548646B (en) | Method for rapidly propagating aralia elata through somatic embryo and secondary somatic embryogenesis | |
| CN103039363B (en) | Rooting medium for tissue culture seedling propagation of camellia oleifera abel and propagation method thereof | |
| CN110384044B (en) | Cultivation method of virus-free seed stems of taros | |
| CN112119913A (en) | Virus-free propagation method for sweet potatoes | |
| CN103477976A (en) | Stem tissue culture seedling method of dendrobium candidum | |
| CN103039362A (en) | Subculture medium for tissue culture seedling propagation of camellia oleifera abel and propagation method thereof | |
| CN107223571B (en) | The quick breeding method for tissue culture in the lobus cardiacus Japanese ardisia | |
| CN103430822B (en) | Aquaculture seed reproduction method for micro seed tubers of konjac | |
| CN100391333C (en) | Aseptic seedling tissue culturing and test tube seedling hardening off and transplating technology for anthurium andraeanum | |
| CN103718963B (en) | A kind of cultural method of horseradish detoxic seedling | |
| CN103858768A (en) | Tissue culture method of plumeria rubra L.cv.Acutifolia | |
| CN111316900A (en) | Method for improving transplanting survival rate of virus-free seedlings of purple-meat sweet potatoes | |
| CN110301335A (en) | The cultivation of the lateral bud breeding bulb of west safflower |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20151218 Address after: 628019 Sichuan Province, Guangyuan City District Road No. 4 Ju Taijie auto service center two building Office District No. 208 Patentee after: Guangyuan Xi Fu Biological Science and Technology Co.,Ltd. Address before: 610106 Sichuan province Chengdu City Shiling Town East Patentee before: Chengdu University |
|
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20150729 |