CN1318578C - Domestication method for lotus root group cultivating de-poisoning breeding - Google Patents
Domestication method for lotus root group cultivating de-poisoning breeding Download PDFInfo
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- CN1318578C CN1318578C CNB2004100653339A CN200410065333A CN1318578C CN 1318578 C CN1318578 C CN 1318578C CN B2004100653339 A CNB2004100653339 A CN B2004100653339A CN 200410065333 A CN200410065333 A CN 200410065333A CN 1318578 C CN1318578 C CN 1318578C
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Abstract
The present invention relates to an acclimation method of lotus root group cultivated detoxified seedlings, which belongs to the technical field of vegetable seedling production. The acclimation method comprises: lotus root stem tips are taken as explants to inoculate and culture non-bacterial seedlings, and inoculated to a culture medium for relay generation propagation; with no need of root culture of group cultivated seedlings, the culture (detoxified) seedlings are directly transferred into a minimum medium with 1 to 3 times of MS or a B5 minimum medium, charged into a culture medium containing 4 to 15% of sucrose and 0 to 1000 mg/L of penicillin, and treated for 7 to 15 days; the lotus root group cultivated (detoxified) seedlings are soaked and washed with tap water, treated for 2 to 48 hrs with 50 to 5000 mg/L of choline chloride, dipped in 50 to 1000 mg/l of NAA and transplanted into shallow water fields directly with a survival rate of over 90%. The group cultivated (detoxified) seedlings are strong in growth vigor, good in production application effect, and suitable for industrial seedling cultivation and commercial production.
Description
One, technical field
The present invention relates to a kind of acclimation method of lotus rhizome group training detoxic seedling.Belong to the vegetable seedling production technical field, be exclusively used in the simple and efficient domestication of lotus rhizome tissue cultured seedling, help the commercial application of lotus rhizome group training (detoxification) seedling.
Two, technical background
Lotus rhizome (Nelumbo nucifera Gaertn) is the traditional characteristics aquatic vegetable of China's southern area, has good nutrient health-care function, also is the main foreign exchange earning vegetables of China.Adopt vegetative propagation to make the virus accumulation because of long-term, each producing region all seriously takes place therefore to cause " stiff lotus root " problem at present, causes kind of a property serious degradation, root stock (lotus root) diminishes, and is stiff, and the product qualitative change is bad, output seriously descends, and has influenced foreign exchange earning and the competitive power on the market at home thereof.Effort improves lotus rhizome quality and output, improves breeding coefficient, and solving and producing " stiff lotus root " problem of going up at present is the task of top priority.Generally adopt annual method of all carrying out strict land for growing field crops seed selection to improve or alleviate the generation of the problems referred to above at present, but labour intensity is big, the technical requirements height, effect is not remarkable, can not really thoroughly solve above-mentioned practical problems.
Present many asexually propagated crops all use shoot tip meristem to cultivate detoxic seedling, and fast numerous virus-elimination seedlings is used production.This maturation of research hexyl of lotus rhizome stem apex detoxify fast breeding technique, but the simple and easy acclimatization technology of lotus rhizome tissue cultured seedling do not appear in the newspapers as yet, influenced the production application of lotus rhizome group training (detoxification) seedling.
Three, summary of the invention
Technical problem
The acclimation method that the purpose of this invention is to provide a kind of lotus rhizome group training virus-elimination seedlings, preferred plan is proposed, root culture during the simplification tissue cultured seedling is used, domestication process and domestication are to the requirement of facility, shortened the acclimation period of lotus rhizome group training (detoxification) seedling, group training (detoxification) seedling growth potential that domestication survives is prosperous, be fit to industrial seedling rearing, be suitable for the land for growing field crops production application.
Technical scheme
The cultural method embodiment of a kind of lotus rhizome group training detoxic seedling provided by the present invention is as follows:
1) getting the lotus rhizome stem apex is explant, is seeded in additional after stem apex outsourcing leaf sheath is divested: MS or the B of 0.01-8.0mg/l6-BA or ZRs and 0.1-10mg/l NAA
5Obtain aseptic seedling in the substratum; After transfer in additional 0.01-10mg/l 6-BA, 0.1-8.0mg/l NAA, 30-50g/l sucrose, the MS of pH5.6-7.2 or B
5Subculture propagation in the substratum,
2) domestication in vitro: tissue cultured seedling is cultivated detoxic seedling with lotus rhizome and is changed 1-3 times of MS or B over to after propagation
5Minimum medium in the substratum of additional 4%-15% sucrose, 100-1000mg/l penicillin, is handled after 7-15 days, and enter and transplant the domestication stage,
In the said process, pH5.6-7.2, intensity of illumination 1800-2500lx, light application time 10-14h/d;
1.3) test tube transplants domestication outward: will after the tap water soaking flushing, handle 2 hours-2 days through the lotus rhizome group training detoxic seedling after domestication training in vitro is strong, directly transplant in the shallow water land for growing field crops after dipping 50-1000mg/LNAA with the 50-5000mg/l choline chloride 60.
Beneficial effect
The present invention compared with prior art has following advantage:
1. compare with planting of present employing with ripe rhizome (lotus root) work, the present invention utilizes the lotus rhizome stem apex to be explant, set up the fast traditional font of lotus rhizome group training (detoxification) system, breeding coefficient reaches 8-20, can obtain a large amount of detoxic seedlings, and be difficult for morphing, group training (detoxification) transplantation of seedlings surviving rate reaches more than 90%, growth potential is prosperous, can significantly reduce sowing quantity, is convenient to the seedling transportation, thereby it is low to have solved lotus rhizome sapling multiplication coefficient of the prior art, sowing quantity is big, transportation and production cost height are unfavorable for the problem that the rapid large-area of improved seeds is applied, and use " stiff lotus root " problem that virus-elimination seedlings can solve serious generation in the present production simultaneously.With generally adopt at present since vegetative period promptly until plant lotus root the selecting and remain for a long time of continuing to carry out of gathering and kind compare, the present invention is by cultivation group training (detoxification) seedling, labour intensity is little, the time length is short, detoxification efficiency is good.
2. the method for lotus rhizome group training (detoxification) seedling simple and efficient provided by the present invention domestication has been omitted tissue cultured seedling and has been used the preceding root culture stage, need not conventional tissue cultured seedling tames desired greenhouse, shades and facility condition and higher administrative skill such as atomizing, the domestication cost is low, and the producer is acceptant.
3. the method for lotus rhizome group training (detoxification) seedling simple and efficient provided by the present invention domestication is applicable to group training (detoxification) seedling of various lotus root lotus kinds.
Four, embodiment
Embodiment 1:
1) acquisition of group training detoxification test tube plantlet: experimental cultivar is " beauty is red " (Nelumbo muciferaGaertn).Getting " beauty is red " stem apex is explant, after the tap water flushing, with 70%7 pure surface sterilizations 1 minute, divide with 0.1% mercuric chloride solution sterilization 15-20 again and plant, aseptic water washing 4-5 all over after, peel off the outsourcing leaf sheath, be inoculated in the triangular flask after cutting stem apex (vegetative point), with MS is minimum medium, and additional 3.0mg/l6-BA and 4.0mg/lNAA obtain aseptic (detoxification) seedling.Above-mentioned test-tube plantlet is cut into band joint individuality be inoculated in the MS minimum medium, additional 2.0mg/l 6-BA, 1.0mg/l NAA carries out shoot proliferation in the MS substratum of 30g/l sucrose.
2) domestication in vitro: lotus rhizome is cultivated (detoxification) seedling change 1.5 times of B over to
5Minimum medium in the substratum of additional 6% sucrose, 300mg/l penicillin, is handled after 12 days, and enter and transplant the domestication stage,
In the said process, pH5.6-7.2, intensity of illumination 1800-2500lx, light application time 10-14h/d;
3) transplant (test tube is outer) domestication: will after the tap water soaking flushing, handle 1 day through lotus rhizome group training (detoxification) seedling after domestication training in vitro is strong, directly transplant in the shallow water land for growing field crops after dipping 200mg/LNAA with the 200mg/L choline chloride 60.Surviving rate can reach 93.3% after 8 days.
Embodiment 2:
1) acquisition of detoxification test tube plantlet: experimental cultivar is " raising lotus root No. one " (Nelumbo nucifera Gaertn) and " beauty is red ".Getting stem apex (or vegetative point) is explant, after the tap water flushing, with 1 fen kind of 70% ethanol surface sterilization, divide kind with 0.1% mercuric chloride solution sterilization 15-20 again, aseptic water washing 4-5 all over after, peel off the outsourcing leaf sheath, be inoculated in the triangular flask after cutting stem apex, with MS is minimum medium, and additional 2.0mg/l ZRs and 1.0mg/l NAA obtain the cultivation seedling.Above-mentioned test-tube plantlet is cut into band joint individuality, be inoculated in B
5In the minimum medium, additional 8.0mg/l ZRs, 2.0mg/l NAA, the B of 50g/l sucrose
5Carry out shoot proliferation in the substratum,
2) domestication in vitro: change lotus rhizome group training (detoxification) seedling over to 2 times of MS minimum mediums, in the substratum of additional 10% sucrose, 500mg/l penicillin, handle after 6 days, enter and transplant the domestication stage,
In the said process, pH5.6-7.2, intensity of illumination 1800-2500lx, light application time 10-14h/d;
3) transplant (test tube is outer) domestication: will after the tap water soaking flushing, handle 12 hours through lotus rhizome group training (detoxification) seedling after the domestication in vitro, directly transplant in the shallow water land for growing field crops after dipping 100mg/lNAA with the 800mg/l choline chloride 60.Surviving rate can reach 95.2% after 5 days.
For purplish red greatly, getting stem apex is explant for the examination lotus root varieties, and according to above method, result of implementation is as follows behind the adjustment substratum composition:
1) test-tube plantlet substratum: MS+2.0mg/l 6-BA+0.5mg/l NAA: shoot proliferation substratum: MS+3.2mg/l 6-BA+2.0mg/l NAA, 30g/l sucrose; Domestication approach: lotus rhizome is cultivated (detoxification) seedling change 3 times of B over to
5Minimum medium, in the substratum of additional 12% sucrose, 200mg/l penicillin, handle after 6 days, lotus rhizome group training (detoxification) seedling is taken out, after the tap water soaking flushing, handled 2 hours with the 5000mg/L choline chloride 60, directly transplant in the shallow water land for growing field crops after dipping 100mg/LNAA, surviving rate can reach 91.2%.
2) test-tube plantlet substratum: MS+3.0mg/l 6-BA+2.0mg/l NAA; Shoot proliferation substratum: MS+5.0mg/l 6-BA+3mg/l NAA, 30g/l white sugar; Domestication approach: lotus rhizome is cultivated (detoxification) seedling change 3 times of B over to
5Minimum medium, in the substratum of additional 8% sucrose, 400mg/l penicillin, handle after 8 days, lotus rhizome group training (detoxification) seedling is taken out, after the tap water soaking flushing, handled 8 hours with the 1000mg/L choline chloride 60, directly transplant in the shallow water land for growing field crops after dipping 500mg/l NAA, surviving rate can reach 92%.
3) test-tube plantlet substratum: MS+6.0mg/l 6-BA+1.0mg/l NAA; Shoot proliferation substratum: MS+6.0mg/l 6-BA+0.5mg/l NAA, 30g/l white sugar; Domestication approach: lotus rhizome is cultivated (detoxification) seedling change 2 times of MS minimum mediums over to, in the substratum of additional 6% sucrose, 500mg/l penicillin, handle after 12 days, lotus rhizome group training (detoxification) seedling is taken out, after the tap water soaking flushing, handled 5 hours with the 2000mg/L choline chloride 60, directly transplant in the shallow water land for growing field crops after dipping 800mg/l NAA, surviving rate can reach 96%.
4) test-tube plantlet substratum: B
5+ 1.0mg/l ZRs+0.5mg/l NAA; Shoot proliferation substratum: B
5+ 2.0mg/lZRs+0.2mg/l NAA, 40g/l white sugar; Domestication approach: change lotus rhizome group training (detoxification) seedling over to 2 times of MS minimum mediums, in the substratum of additional 6% sucrose, 250mg/l penicillin, handle after 12 days, lotus rhizome group training (detoxification) seedling is taken out, after the tap water soaking flushing, handled 5 hours with the 2000mg/L choline chloride 60, directly transplant in the shallow water land for growing field crops after dipping 800mg/l NAA, surviving rate can reach 96%.
5) test-tube plantlet substratum: MS+7.0mg/l ZRs+2.0mg/l NAA; Shoot proliferation substratum: B
5+ 4.0mg/l ZRs+1.0mg/l NAA, 40g/l white sugar; Domestication approach: lotus rhizome is cultivated (detoxification) seedling change 2 times of B over to
5Minimum medium, in the substratum of additional 10% sucrose, 600mg/l penicillin, handle after 6 days, lotus rhizome group training (detoxification) seedling is taken out, after the tap water soaking flushing, handled 8 hours with the 1000mg/L choline chloride 60, directly transplant in the shallow water land for growing field crops after dipping 800mg/l NAA, surviving rate can reach 92.7%.
6) test-tube plantlet substratum: B
5+ 3.0mg/l ZRs+3.0mg/l NAA; Shoot proliferation substratum: B
5+ 6.0mg/lZRs+1.0mg/l NAA, 30g/l sucrose; Domestication approach: lotus rhizome is cultivated (detoxification) seedling change 1.5 times of MS minimum mediums over to, in the substratum of additional 15% sucrose, 1000mg/l penicillin, handle after 5 days, lotus rhizome group training (detoxification) seedling is taken out, after the tap water soaking flushing, handled 8 hours with the 3000mg/L choline chloride 60, directly transplant in the shallow water land for growing field crops surviving rate 91.5% after dipping 1000mg/l NAA.
7) test-tube plantlet substratum: MS+2.0mg/l 6-BA+1.0mg/l NAA; Shoot proliferation substratum: B
5+ 3.0mg/l ZRs+0.5mg/l NAA, 40g/l white sugar; Domestication approach: change lotus rhizome group training (detoxification) seedling over to MS times of minimum medium, in the substratum of additional 12% sucrose, 800mg/l penicillin, handle after 5 days, lotus rhizome group training (detoxification) seedling is taken out, after the tap water soaking flushing, handled 4 hours with the 4000mg/L choline chloride 60, directly transplant in the shallow water land for growing field crops surviving rate 93.5% after dipping 1000mg/l NAA.
8) test-tube plantlet substratum: B
5+ 2.0mg/l 6-BA+0.2mg/l NAA; Shoot proliferation substratum: MS+4.0mg/l ZRs+1.0mg/l NAA, 40g/l white sugar; Domestication approach: change lotus rhizome group training (detoxification) seedling over to 3 times of MS times of minimum mediums, in the substratum of additional 9% sucrose, 600mg/l penicillin, handle after 8 days, lotus rhizome group training (detoxification) seedling is taken out, after the tap water soaking flushing, handled 4 hours with the 3000mg/L choline chloride 60, directly transplant in the shallow water land for growing field crops surviving rate 93.5% after dipping 2000mg/l NAA.
9) test-tube plantlet substratum: MS+7.0mg/l 6-BA+2.0mg/l NAA; Shoot proliferation substratum: MS+5.0mg/l 6-BA+1.0mg/l NAA, 50g/l white sugar; Domestication approach: change lotus rhizome group training (detoxification) seedling over to 3 times of B
5Minimum medium in the substratum of additional 8% sucrose, 500mg/l penicillin, is handled after 8 days, lotus rhizome group training (detoxification) seedling is taken out, after the tap water soaking flushing, handled 10 hours with the 1000mg/L choline chloride 60, directly transplant in the shallow water land for growing field crops surviving rate 95% after dipping 1500mg/l NAA.
10) test-tube plantlet substratum: B
5+ 3.0mg/l ZRs+1.5mg/l NAA; Shoot proliferation substratum: MS+6.0mg/l ZRs+1.5mg/l NAA, 30g/l sucrose; Domestication approach: lotus rhizome is cultivated (detoxification) seedling change 1.5 times of MS minimum mediums over to, in the substratum of additional 10% sucrose, 700mg/l penicillin, handle after 12 days, lotus rhizome group training (detoxification) seedling is taken out, after the tap water soaking flushing, handled 12 hours with the 3000mg/L choline chloride 60, directly transplant in the shallow water land for growing field crops surviving rate 91.6% after dipping 100mg/l NAA
11) test-tube plantlet substratum: B
5+ 2.0mg/l ZRs+0.2mg/l NAA; Shoot proliferation substratum: MS+5.0mg/l ZRs+1.0mg/l NAA, 30g/l sucrose; Domestication approach: lotus rhizome is cultivated (detoxification) seedling change 1.5 times of MS minimum mediums over to, in the substratum of additional 12% sucrose, 500mg/l penicillin, handle after 8 days, lotus rhizome group training (detoxification) seedling is taken out, after the tap water soaking flushing, handled 24 hours with the 800mg/L choline chloride 60, directly transplant in the shallow water land for growing field crops surviving rate 94.6% after dipping 500mg/l NAA.
12) test-tube plantlet substratum: MS+2.0mg/l 6-BA+0.1mg/l NAA; Shoot proliferation substratum: B
5+ 5.0mg/l ZRs, 40g/l white sugar; Domestication approach: lotus rhizome is cultivated (detoxification) seedling change 2.5 times of B over to
5Minimum medium, in the substratum of additional 15% sucrose, 1000mg/l penicillin, handle after 6 days, lotus rhizome group training (detoxification) seedling is taken out, after the tap water soaking flushing, handled 24 hours with the 1500mg/L choline chloride 60, directly transplant in the shallow water land for growing field crops surviving rate 95% after dipping 200mg/l NAA.
13) test-tube plantlet substratum: B
5+ 3.0mg/l 6-BA+1.0mg/l NAA; Shoot proliferation substratum: MS+6.0mg/l ZRs+3.0mg/l NAA, 40g/l sucrose; Domestication approach: change lotus rhizome group training (detoxification) seedling over to 3 times of B
5Minimum medium, in the substratum of additional 10% sucrose, 800mg/l penicillin, handle after 6 days, lotus rhizome group training (detoxification) seedling is taken out, after the tap water soaking flushing, handled 12 hours with the 3000mg/L choline chloride 60, directly transplant in the shallow water land for growing field crops surviving rate 92.5% after dipping 1500mg/l NAA.
14) test-tube plantlet substratum: MS+5.0mg/l 6-BA+2.0mg/l NAA; Shoot proliferation substratum: B
5+ 4.0mg/l ZRs+0.1mg/l NAA; Domestication approach: lotus rhizome is cultivated (detoxification) seedling change 3 times of B over to
5Minimum medium, in the substratum of additional 10% sucrose, 600mg/l penicillin, handle after 6 days, lotus rhizome group training (detoxification) seedling is taken out, after the tap water soaking flushing, handled 12 hours with the 200mg/L choline chloride 60, directly transplant in the shallow water land for growing field crops surviving rate 90.5% after dipping 1500mg/l NAA.
15) test-tube plantlet substratum: MS+3.0mg/l 6-BA+0.5mg/l NAA; Shoot proliferation substratum: MS+3.0mg/l ZRs+0.1mg/l NAA, 30g/l sucrose; Domestication approach: change lotus rhizome group training (detoxification) seedling over to 2 times of MS minimum mediums, in the substratum of additional 9% sucrose, 300mg/l penicillin, handle after 8 days, lotus rhizome group training (detoxification) seedling is taken out, after the tap water soaking flushing, handled 24 hours with the 200mg/L choline chloride 60, directly transplant in the shallow water land for growing field crops surviving rate 95% after dipping 1500mg/l NAA.
16) test-tube plantlet substratum: B
5+ 3.0mg/l ZRs+2.0mg/l NAA; Shoot proliferation substratum: B
5+ 6.0mg/l ZRs+0.5mg/l NAA, 50g/l sucrose; Domestication approach: lotus rhizome is cultivated (detoxification) seedling change 2 times of MS minimum mediums over to, in the substratum of additional 6% sucrose, 100mg/l penicillin, handle after 12 days, lotus rhizome group training (detoxification) seedling is taken out, after the tap water soaking flushing, handled 24 hours with the 200mg/L choline chloride 60, directly transplant in the shallow water land for growing field crops surviving rate 91.5% after dipping 500mg/l NAA.
17) test-tube plantlet substratum: MS+8.0mg/l 6-BA+2.0mg/l NAA; Shoot proliferation substratum :+6.0mg/l ZRs+1.0mg/l NAA, 50g/l sucrose; Domestication approach: lotus rhizome is cultivated (detoxification) seedling change 3 times of B over to
5Minimum medium in the substratum of additional 6% sucrose, 200mg/l penicillin, is handled after 15 days, lotus rhizome group training (detoxification) seedling is taken out, after the tap water soaking flushing, handled 24 hours with the 500mg/L choline chloride 60, directly transplant in the shallow water land for growing field crops surviving rate 93% after dipping 500mg/l NAA.
18) test-tube plantlet substratum: B
5+ 3.0mg/l ZRs+1.0mg/l NAA; Shoot proliferation substratum: B
5+ 6.0mg/l ZRs+0.5mg/l NAA, 40g/l sucrose; Domestication approach: lotus rhizome is cultivated (detoxification) seedling change 1.5 times of MS minimum mediums over to, in the substratum of additional 6% sucrose, 1000mg/l penicillin, handle after 12 days, lotus rhizome group training (detoxification) seedling is taken out, after the tap water soaking flushing, handled 24 hours with the 2000mg/L choline chloride 60, directly transplant in the shallow water land for growing field crops surviving rate 96.5% after dipping 1000mg/l NAA.
19) test-tube plantlet substratum: MS+8.0mg/l ZRs+2.0mg/l NAA; Shoot proliferation substratum: B
5+ 6.0mg/l ZRs+0.5mg/l NAA, 50g/l sucrose; Domestication approach: lotus rhizome is cultivated (detoxification) seedling change 3 times of B over to
5Minimum medium, in the substratum of additional 15% sucrose, 800mg/l penicillin, handle after 8 days, lotus rhizome group training (detoxification) seedling is taken out, after the tap water soaking flushing, handled 4 hours with the 5000mg/L choline chloride 60, directly transplant in the shallow water land for growing field crops surviving rate 91.5% after dipping 1000mg/l NAA.
Claims (1)
1. the acclimation method of lotus rhizome group training detoxic seedling, implementation step is as follows:
1.1) to get the lotus rhizome stem apex be explant, is seeded in additional after stem apex outsourcing leaf sheath is divested: MS or the B of 0.01-8.0mg/l 6-BA or ZRs and 0.1-10mg/l NAA
5Substratum obtains aseptic seedling; After transfer in additional 0.01-10mg/l 6-BA, 0.1-8.0mg/l NAA, the MS of 30-50g/l sucrose or B
5Subculture propagation in the substratum,
1.2) domestication in vitro: tissue cultured seedling is cultivated detoxic seedling with lotus rhizome and is changed 1-3 times of MS or B over to after propagation
5Minimum medium in the substratum of additional 4%-15% sucrose, 100-1000mg/l penicillin, is handled after 7-15 days, and enter and transplant the domestication stage,
In the said process, pH5.6-7.2, intensity of illumination 1800-2500lx, light application time 10-14h/d;
1.3) test tube transplants domestication outward: will after the tap water soaking flushing, handle 2 hours-2 days through the lotus rhizome group training detoxic seedling after domestication training in vitro is strong, directly transplant in the shallow water land for growing field crops after dipping 50-1000mg/LNAA with the 50-5000mg/l choline chloride 60.
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| CN104542297A (en) * | 2015-01-23 | 2015-04-29 | 杨学 | Cultivation method of nine-grade nymphaea hybrid |
| CN105325298B (en) * | 2015-11-18 | 2017-12-22 | 扬州大学 | A kind of lotus rhizome stem-tip tissue Initial culture method |
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Non-Patent Citations (4)
| Title |
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| 氯化胆碱对大岩桐组织陪苗生长的影响 何若天等，广西农业生物科学，第21卷第2期 2002 * |
| 莲耦组织培养及快速繁殖试验研究 于文进等，广西热作科技，第2期 1999 * |
| 莲耦茎尖培养技术的初步研究 李良俊等，江苏农学院学报，第16卷第3期 1995 * |
| 莲耦茎尖培养技术的初步研究 李良俊等，江苏农学院学报，第16卷第3期 1995;莲耦组织培养及快速繁殖试验研究 于文进等，广西热作科技，第2期 1999;氯化胆碱对大岩桐组织陪苗生长的影响 何若天等，广西农业生物科学，第21卷第2期 2002 * |
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