CN103070052B - Method for breeding regeneration plants by rice dormant buds - Google Patents
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Abstract
The invention discloses a method for breeding regeneration plants by rice dormant buds, and belongs to the technology and bleeding field of crop seedling. According to the method, rice straws of rice with reciprocal 1 to reciprocal 3 sections of dormant buds are taken to be scattered and planted onto a seedling culture bed, soil is covered, or the rice straws are soaked by plant hormone, then, the rice straws are scattered and planted onto the seedling culture bed, the soil is covered, the seedling culture and the management are carried out according to the conventional seedling culture method, and the seedlings are transplanted to the field after the regeneration seedlings reach 3-leaf to 4-leaf period. The method has the advantages that the effective storage and breeding can be realized for obtaining a large number of special rice materials including female sterile, male sterile and the like, the difficult problem of breeding difficulty of special materials is solved, hybrid F1 can be directly utilized for obtaining a large number of F1 regeneration plants, the rice hybrid seed production cost can be effectively reduced, the breeding period is shortened, and the hybrid advantages can be perfectly maintained. According to the method, the rice overground parts with rice straws with reciprocal 1 to reciprocal 3 sections of dormant buds are adopted, both the regeneration efficiency of the rice straws treated by hormone and the regeneration efficiency of the rice straws which are not treated by hormone are obviously superior to that of rice pile direct regeneration, the method is simple, the implementation is easy, and the popularization and the application are convenient.
Description
Technical field
The present invention relates to a kind of method of utilizing paddy rice sleeping bud to obtain regeneration plant, belong to agricultural biological technical field, more particularly belong to plant breeding technique field.
Background technology
Paddy rice (
oryza sativa.l) be the third place in the world generalized grain crop that is only second to corn and wheat, having in the world population over half is to take rice as staple food.China is world paddy rice production and consumption big country, and rice cropping area accounts for 1/3 of plant of grain crops area, and output accounts for the nearly 1/2 of total output of grain, and per capita year more than consumption rice 150kg, so Rice Production is concerning the safe overall situation of national food.Constantly perfect along with modern Rice Production technology all obtained achievement highly visible in the raising of yield of brown rice and quality, alleviated greatly the pressure of crisis in food.But since 20 end of the centurys, population in the world is the situation in rapid growth always, causes and start now to occur that consumption figure is greater than output, and stockpile number atrophy, price are unprecedented soaring.Therefore in the face of so severe population problem, grain-production shoulders heavy responsibilities.
Ensure grain yield steady-state growth, just need constantly to excavate by all means new germ plasm resource, open up new production technology, seek economy and efficient production model.Therefore realize this goal, just need on the production chain of paddy rice, obtain new breakthrough.Discovery and the further investigation of the paddy female sterile mutant material of the exploitation of Exploration of Wild Rice Germplasm Resources, the spread of ratoon rice, Dominant gene, and take this series of research such as new crossbreeding pattern of seeking all as we provide good thinking of development.
The breeding of paddy rice also depends on sexual propagation and vegetative propagation as other plant.Sexual propagation, as its topmost modes of reproduction, all plays decisive role to the production of paddy rice and breeding.Utilization and the development of male sterility of rice (male sterility, MS) and paddy female sterile (femalesterility, FS), hybrid rice is the Important Action of Ensuring Food Safety.At present, in breeding of hybridized rice, development and the technical system of utilizing mainly contain three line method and two line method, although these systems are all comparatively ripe, but remain some shortcomings, be mainly reflected in following some: 1. Parent branch or subregion plantation, while blooming, must manually catch up with powder supplementary pollination, during results, Parent people work point is received, time-consuming taking a lot of work; 2. hybrid seed yield is low, can not fine solution impurity of seeds problem; 3. be not suitable for scale, mechanized operation etc.Based on this, visible crossbreeding process also needs to expend huge material resources, manpower and very long breeding cycle, could seek so a kind of with F
1for material, by the means of vegetative propagation technique, obtain fast the production of hybrid seeds pattern of large batch of hybrid strain? along with the application of paddy female sterile in breeding, utilize male sterile (MS) and female sterile (FS), build the production of hybrid seeds pattern of " MS+FS ", pollen donor while being the production of hybrid seeds is Female sterile clone, and being also expected to soon follow-up three is to carry out large-scale promotion and application with the another Breeding Model of double-line hybrid.But it is Female sterile clone that problem is its male parent, can not be normally solid to obtain large batch of female sterile strain.Therefore, can the application of succeeding of this pattern, depend on and obtain a large amount of female sterile strains.Also have some special rare your materials (as monomer and trisome and polyploid, the material such as selfing is not affine), because breeding be bred or be difficult to the feature of self can not by amphigenetic mode.Vegetative propagation technique and gene engineering etc. are how to obtain the most frequently used method and approach of these special materials at present.
The vegetative propagation of plant mainly contains cuttage, grafting, press strip and tissue cultivation etc., to organize, cultivates being most widely used in paddy rice.But cultivating, rice tissue also there is larger limitation: it draws materials rataria, young fringe, pollen and be subject to season and environmental limit, and the time of drawing materials is strict, too early or spend evening and all can cause callus of induce failure; In tissue culture procedures, easily there is the situations such as death, albefaction, be difficult to obtain a large amount of group training seedlings, and group training seedling relatively a little less than, be transplanted to land for growing field crops survival rate and can not get ensureing, the while is high for large-scale production cost.
Wild rice is made the important materials of excavating resource as recent rice, be more and more subject to researcher's attention.Common wild-rice is the nearly edge ancestors of cultivated rice, and it,, through evolution all the year round, becomes modern cultivated rice.Cultivated rice is generally annual, and common wild-rice is herbaceos perennial, has stronger reviviscence.Utilize ratoon rice that this characteristic grows up now to have the application of certain scale, obtained appreciable achievement.At present, approximately 1,100 ten thousand mu of China's ratoon rice cultivated areas, are the countries that develops in the world ratoon rice area maximum.But ratoon rice plant height, tiller, the proterties such as spike length, Number of kernels compared first rice crop and all relatively fallen sharply, and causes average unit yield not high, output is unstable.At present rice regeneration strain, only for directly utilize rice stub regeneration in field, does not only make full use of its sleeping bud regeneration capacity, and output and productivity effect also generally on the low side.Utilize the reproducing characteristic of paddy rice sleeping bud to breed aftergrowth, also there is no a set of production technology system that can obtain fast regeneration strain.Reasonably excavate paddy rice sleeping bud regeneration capacity, in conjunction with plant hormone and the regeneration of growth regulator induction paddy rice sleeping bud, obtain regeneration plant and build perfect culture technique system, the research of this respect need further deeply.
Summary of the invention
The technical problem to be solved in the present invention is to overcome existing rice regeneration strain only for directly utilize rice stub regeneration in field, output is unstable, output and productivity effect are generally on the low side, and the production cost of hybrid rice is high and production of hybrid seeds process is loaded down with trivial details, the breeding difficulty of special material, paddy rice group training seedling easily occurs that a little less than death, albefaction and group training seedling are relatively, being transplanted to land for growing field crops survival rate can not get ensureing, there is defect and deficiency that large-scale production cost is high, the object of this invention is to provide efficient, quick, the low-cost paddy rice mating system that obtains rice regeneration strain of a kind of energy.
Paddy rice sleeping bud provided by the present invention is bred the method for regeneration plant, comprises the following steps:
(1) at first rice crop, gathered in latter 7 days, intercepting first rice crop acrial part with falling 1～fall the rice straw of 3 node dormancy buds;
(2) be spread out on seedling bed and earthing after rice straw step (1) Suo Shu is cleaned, by the conventional seedbed of paddy rice method of raising seedling, grow seedlings and manage, or after being cleaned, rice straw step (1) Suo Shu soaks 40～50 hours with plant hormone composition, be spread out over again on seedling bed and earthing, by the conventional seedbed of paddy rice method of raising seedling, grow seedlings and manage;
(3) treat regrowth 3 leaf～4 leaf phases transplanting land for growing field crops, by the conventional field production management method of paddy rice, carry out field production management.
The formula of described plant hormone composition is 0.25mg/L 6-BA+30 mg/L～60mg/L NAA, 0.1mg/L NAA+10.0 mg/L～12.0mg/L 6-BA or 10mg/L Kinetin+2.0 mg/L～5.0mg/L NAA.
The formula of described plant hormone composition is 0.25mg/L 6-BA+30mg/L NAA.
One of feature of the present invention is the sleeping bud utilizing on paddy rice rice straw node, builds the technical system that obtains fast a large amount of rice regeneration plant.Sleeping bud also claims axillalry bud, resting bud or regeneration bud, is the basis of rice regeneration.
Second feature of the present invention is in conjunction with plant hormone induction, improves Dormant Buds efficiency and promotes growing of regrowth.Plant hormone (Plant hormone), is that some are synthetic in plant corpus, can be delivered to site of action from producing position, and micro-concentrations can produce to plant corpus the acitve organic matter of certain physiological action.It can also be the similar chemical substance (plant growth regulator) of artificial composite structure effect.Many plant hormones, on the regeneration of paddy rice and the impact of having grown, have the Cell Differentiation of promotion, division, growth, and the dormancy of breaking dormancy bud regulates the transportation of nutriment, is also beneficial to the effects such as induction root and adventive root formation simultaneously.
Compared with prior art, the invention has the beneficial effects as follows:
(1) can effectively preserve and breed and obtain a large amount of special rice material (as female sterile, male sterile, selfing are not affine, monomer and polyploid etc.), solve the difficult problem that special material is difficult for breeding.
(2) directly utilize hybrid F
1just can obtain a large amount of F
1regeneration plant, does not need to carry out hybrid seeding repeatedly, not only effectively reduces the cost of the paddy rice cross breeding production of hybrid seeds and shortens breeding cycle, can also well keep its hybrid vigour.
(3) because the regeneration capacity of different materials is different, in conjunction with plant hormone, use the germination efficiency of the sleeping bud that can improve the relatively poor kind of regeneration power and the growth that promotes node new root, well strengthened the scope of application of the method.
(4) asexual reproduction method that the present invention sets up, for special rice material (educate as female, female sterile, male sterile etc.) and conventional Rice, hybrid rice (hybrid F
1) adopt with falling 1～fall 3 node dormancy buds rice straw with HORMONE TREATMENT with need not hormone-treated regeneration efficiency all extremely significantly be better than using rice stub Direct Regeneration, its emergence rate, leaf area index, plant height, Number of kernels geometric ratio contrast and increase substantially, difference is extremely remarkable.Need not hormone-treated method, it is emerged efficiency and improves 68.7%～91.2% than the efficiency of emerging of directly transplanting rice stub, and Number of kernels is than the Number of kernels improve 105.5%～158.2% of directly transplanting rice stub; Use hormone-treated method, its emergence rate has improved 112.3%～132.9% than the emergence rate of the contrast of direct transplanting rice pile material, and grain husk spends number to improve 107.1%～138.3% than the contrast of direct transplanting rice pile material, has all produced unforeseeable technique effect.
(5) the inventive method simply and is easily implemented, easy to utilize.
Accompanying drawing explanation
Fig. 1 is the emergence rate statistical chart that first round hormon, different hormone combinations thing and variable concentrations are processed rice material, and in figure, abscissa represents the concentration of hormone and hormonal composition, and ordinate represents the emergence rate of rice material, curve in Figure 1A
represent emergence rate, curve that rice material is processed with variable concentrations NAA
represent emergence rate, curve that rice material is processed with variable concentrations KIN
represent emergence rate, curve that rice material is processed with variable concentrations 6-BA
represent the emergence rate that rice material is processed with KN hormonal composition listed in table 2; Curve in Figure 1B
represent the emergence rate that rice material is processed with DB hormonal composition listed in table 2, curve
represent the emergence rate that rice material is processed with NB hormonal composition listed in table 2, curve
represent the emergence rate that rice material is processed with DK hormonal composition listed in table 2, curve
represent the emergence rate that rice material is processed with NK hormonal composition listed in table 2.
Fig. 2 second takes turns the emergence rate statistical chart of processing rice material with DK hormonal composition variable concentrations, and in figure, abscissa represents the concentration of DK hormonal composition, and ordinate represents to process the emergence rate of rice material, curve
represent the emergence rate that fs processes by hormon concentration listed in table 3, curve
represent the emergence rate that FS processes by hormon concentration listed in table 3, curve
represent the emergence rate that CMS processes by hormon concentration listed in table 3, curve
represent the emergence rate that multitude elm B processes by hormon concentration listed in table 3, curve
represent the emergence rate that in the assorted 37 use tables 3 in Yunnan, listed hormon concentration is processed.
Fig. 3 second takes turns the emergence rate statistical chart of processing rice material with NB hormonal composition variable concentrations, and in figure, abscissa represents the concentration of NB hormonal composition, and ordinate represents to process the emergence rate of rice material, curve
represent the emergence rate that fs processes by hormon concentration listed in table 4, curve
represent the emergence rate that FS processes by hormon concentration listed in table 4, curve
represent the emergence rate that CMS processes by hormon concentration listed in table 4, curve
represent the emergence rate that multitude elm B processes by hormon concentration listed in table 4, curve
represent the emergence rate that in the assorted 37 use tables 4 in Yunnan, listed hormon concentration is processed.
Fig. 4 second takes turns the emergence rate statistical chart of processing rice material with BN hormonal composition variable concentrations, and in figure, abscissa represents the concentration of BN hormonal composition, and ordinate represents the emergence rate of rice material, curve
represent the emergence rate that fs processes by hormon concentration listed in table 5, curve
represent the emergence rate that FS processes by hormon concentration listed in table 5, curve
represent the emergence rate that CMS processes by hormon concentration listed in table 5, curve
represent the emergence rate that multitude elm B processes by hormon concentration listed in table 5, curve
represent the emergence rate that in the assorted 37 use tables 5 in Yunnan, listed hormon concentration is processed.
Fig. 5 second takes turns the emergence rate statistical chart of processing rice material with KN hormonal composition variable concentrations, and in figure, abscissa represents the concentration of KN hormonal composition, and ordinate represents the emergence rate of rice material, curve
represent the emergence rate that fs processes by hormon concentration listed in table 6, curve
represent the emergence rate that FS processes by hormon concentration listed in table 6, curve
represent the emergence rate that CMS processes by hormon concentration listed in table 6, curve
represent the emergence rate that multitude elm B processes by hormon concentration listed in table 6, curve
represent the emergence rate that in the assorted 37 use tables 6 in Yunnan, listed hormon concentration is processed.
Embodiment is to disseminate and cultivate regeneration or regenerate in conjunction with plant hormone induction culturing with vegetative approach with the sleeping bud of first rice crop rice straw Nodes, the breeding of regenerating of the final method that obtains recycled water rice plants, the rice stub of take seed sprouting, directly transplanting is contrast, relatively their difference on each economical character.
Following case study on implementation all arranges three and repeats above experiment; Experiment material used and reagent all can obtain in business purchase.
embodiment 1
Select FS (female educating), fs(female sterile), CMS(cytoplasmic male sterility cytoplasmic male sterility), multitude elm B(conventional material), the assorted 37(hybrid F in Yunnan
1) be test material, by disseminate earthing with the rice straw of node dormancy bud, cultivate, and direct transplanting rice stub is set for contrast, to emergence rate in seedling stage, maturing stage is carried out the investigation such as plant height, spike length, Number of kernels, 100-grain weight, investigates the inventive method to advantages such as efficiency of emerging such as some paddy rice special materials, conventional material, hybrid rices.
Concrete grammar is as follows:
(1) at FS, fs, CMS, multitude elm B, assorted 37 first rice crops in Yunnan, gather in latter 7 days, intercepting FS, fs, CMS, multitude elm B, the assorted 37 first rice crop acrial parts in Yunnan with falling 1～fall the rice straw of 3 node dormancy buds, clean, (described first rice crop acrial part with falling 1～specifically the drawing materials of rice straw of 3 node dormancy buds of falling is to get the above approximately rice straw of 10～15cm in earth's surface, it is with falling 1 and fall the rice straw of two node dormancy buds of 2 nodes, or with falling 1, fall 2 and fall the rice straw of three node dormancy buds of 3 nodes, consider practical condition, this test by above-mentioned with falling 1 and fall two node dormancy buds of 2 node and with falling 1, fall 2 and fall the same material of three node dormancy buds of 3 nodes and be mixed and grow seedlings, following embodiment with falling 1～fall 3 node dormancy buds rice straw specifically draw materials all identical with the present embodiment),
(2) the above-mentioned rice straw with node dormancy bud is evenly spread out over to earthing on seedling bed by same material Yi Ge community, by the conventional seedbed of paddy rice method of raising seedling, grow seedlings and manage, transplant FS rice stub, fs rice stub, CMS rice stub, multitude elm B rice stub, assorted 37 rice stubs in Yunnan to identical seedling bed simultaneously, by the conventional seedbed of paddy rice method of raising seedling, grow seedlings and manage, in contrast;
(3) by process and the regrowth of contrast in the 3 leaf-4 leaf phases statistical comparison efficiency of emerging respectively, and Transplanting Test field, carries out field production management by the conventional field production management method of paddy rice.
Result of the test is found (table 1): utilize the inventive method by with falling 1～that the rice straw of 3 node dormancy buds of falling is disseminated the efficiency of emerging in seedling stage that earthing cultivates is more much higher than the efficiency of emerging of directly transplanting rice stub, its emergence rate: FS improves 68.7% than contrast, fs improves 73.8% than contrast, CMS improves 80.4% than contrast, multitude elm B improves 80.3% than contrast, and Yunnan assorted 37 improves 91.2% than contrast; Its Number of kernels (grain/fringe): FS improves the contrast of 158.2%, fs ratio than contrast and improves 146.6%, CMS than contrast raising 146.9%, and multitude elm B improves 108.6% than contrast, and the assorted 37 ratio contrasts in Yunnan improve 105.5%.The rice straw that is about to falling 1～falls 3 node dormancy buds is disseminated earthing and is cultivated, it emerges efficiency than the efficiency raising 68.7%～91.2% of emerging of directly transplanting rice stub, its Number of kernels is than the Number of kernels improve 105.5%～158.2% of directly transplanting rice stub, shows that the inventive method is for special rice material (educate as female, female sterile, male sterile etc.) and conventional Rice, hybrid F
1employing with falling 1～rice straw of 3 node dormancy buds of falling breeds acquisition regeneration plant, its emergence rate, Number of kernels and respective material are directly transplanted rice stub and are compared and all produced unforeseeable technique effect.
embodiment 2
Selecting FS, fs, CMS, multitude elm B, Yunnan assorted 37 is test material, utilize the KIN(kinetin of different gradients), 6-BA(6-benzyladenine), NAA (methyl α-naphthyl acetate)+KIN, 2,4-D(2,4-dichlorphenoxyacetic acid)+KIN, NAA+6-BA, 2,4-D+6-BA solution concentration is processed, the projects such as emergence rate are investigated, compare the situations such as emergence rate under hormon, different hormone combinations and variable concentrations, through the screening of number wheel, obtain suitable hormone, hormone combinations and concentration ratio.
Concrete grammar is as described below:
(1) at FS, fs, CMS, multitude elm B, assorted 37 first rice crops in Yunnan, gather in latter 7 days, intercepting FS, fs, CMS, multitude elm B, the assorted 37 first rice crop acrial parts in Yunnan with falling 1～fall the rice straw of 3 node dormancy buds, clean;
(2) rice straw in FS, the fs described in mixed in equal amounts step (1), CMS, multitude elm B, Yunnan assorted 37, soaks 48h processing with hormone, the hormonal composition of the variable concentrations shown in table 2 respectively;
(3) rice straw with node dormancy bud after step (2) processing is evenly spread out over to earthing on seedling bed, by the conventional seedbed of paddy rice method of raising seedling, grows seedlings and manage;
(4) treat that the 3 leaf-4 leaf phases of regrowth add up respectively emergence rate, just sift out good hormone, hormonal composition and concentration (see figure 1).
Table 2 annotation:
1, table 2 plant hormone used and the abbreviation of plant hormone composition:
Kinetin（KIN）、6-BA（BA）、0.1mg/LNAA+Kinetin（NK）、0.1mg/L?2,4-D+Kinetin（DK）、0.1mg/L?NAA+6-BA（NB）、0.1mg/L?2,4-D+6-BA（DB）、10mg/L?Kinetin+?NAA（KN）；
2, other symbol of table 2 represents:
KIN1～KIN6, BA1～BA6, NAA1～NAA6 represent that respectively hormone Kinetin, 6-BA, NAA variable concentrations gradient are followed successively by: 1mg/L, 3mg/L, 6mg/L, 9mg/L, 12mg/L, 15mg/L;
NK1～NK6 represents that respectively, in hormonal composition 0.1mg/L NAA+Kinetin, the concentration quantitative of NAA is 0.1mg/L, and in this hormonal composition, the change in concentration gradient of KIN is followed successively by 1mg/L, 3mg/L, 6mg/L, 9mg/L, 12mg/L, 15mg/L;
DK1～DK6 represents respectively hormonal composition 0.1mg/L2, and in 4-D+Kinetin, the concentration quantitative of 2,4-D is 0.1mg/L, and in this hormonal composition, the change in concentration gradient of KIN is followed successively by 1mg/L, 3mg/L, 6mg/L, 9mg/L, 12mg/L, 15mg/L;
NB1～NB6 represents that respectively, in hormonal composition 0.1mg/L NAA+6-BA, the concentration quantitative of NAA is 0.1mg/L, and in this hormonal composition, the change in concentration gradient of 6-BA is followed successively by 1mg/L, 3mg/L, 6mg/L, 9mg/L, 12mg/L, 15mg/L;
DB1～DB6 represents respectively hormonal composition 0.1mg/L 2, and in 4-D+6-BA, the concentration quantitative of 2,4-D is 0.1mg/L, and in this hormonal composition, the change in concentration gradient of 6-BA is followed successively by 1mg/L, 3mg/L, 6mg/L, 9mg/L, 12mg/L, 15mg/L;
KN1～KN6 represents that respectively in hormonal composition 10mg/L Kinetin+ NAA, the concentration quantitative of KIN is 10mg/L, and in this hormonal composition, the change in concentration gradient of NAA is followed successively by 1mg/L, 3mg/L, 6mg/L, 9mg/L, 12mg/L, 15mg/L.
Result is found (Fig. 1):
While using single hormone, along with the raising of hormone concentration, emergence rate is all parabola trend.Use Kinetin emergence rate to rise along with the rising of concentration, be about 36% 10mg/L emergence rate is the highest, decline afterwards; Use 6-BA emergence rate to raise along with the rising of concentration, when concentration is 9mg/L, emergence rate is up to 37%, and the rising emergence rate along with concentration sharply reduces afterwards; Use NAA emergence rate to raise along with the rising of concentration, when concentration is 15mg/L, emergence rate reaches 38%, and the trend increasing in addition along with the rising emergence rate of concentration.
DB(is hormonal composition: 0.1mg/L 2,4-D+6-BA), NK(0.1mg/L NAA+Kinetin) emergence rate is all lower than 40%, induction emergence rate is all undesirable.
KN(10mg/L Kinetin+ NAA) in hormone combinations, take the KIN of 10.0mg/L as quantitatively, the concentration of NAA is higher at 1.0mg/L～6.0mg/L emergence rate, all higher than 40%, has good induction efficiency, concentration along with NAA raises afterwards, and emergence rate obviously reduces.
DK(0.1mg/L2,4-D+Kinetin) in hormone combinations with 2 of 0.1mg/L, 4-D is that quantitatively the concentration of Kinetin is higher at 2.0mg/L～4.0mg/L emergence rate, all higher than 40%, along with the concentration of Kinetin raises, emergence rate obviously reduces.
NB(0.1mg/L NAA+6-BA) in hormone combinations, take the NAA of 0.1mg/L as quantitatively, the concentration of 6-BA is higher at 7.0mg/L～12.0mg/L emergence rate, and all higher than 40%, the concentration along with 6-BA raises afterwards, and emergence rate obviously reduces.
Therefore, the result of primary dcreening operation:
While 1. using a kind of hormone separately, when Kinetin concentration is 10mg/L, emergence rate is up to 36%, and other plant hormone capable of being combined further screens; When NAA concentration is 15mg/L, emergence rate reaches 38%, and the trend increasing in addition along with the rising emergence rate of concentration, can continue the screening of higher concentration; When 6-BA concentration is 1mg/L～10mg/L, emergence rate is all higher than 30%, and when 9mg/L, emergence rate is up to 37%, and other plant hormone capable of being combined further screens (seeing Figure 1A).
While 2. combining Corticosteroids, DK hormonal composition: 0.1mg/L 2,4-D+2.0mg/L～4.0mg/LKinetin,
Its emergence rate is higher, all higher than 40%, can further screen (seeing Figure 1B); NB hormonal composition: 0.1mg/L NAA+7.0mg/L～12.0mg/L 6-BA, emergence rate is higher, all higher than 40%, can further screen (seeing Figure 1B); KN hormonal composition: 10.0mg/L Kinetin+1.0mg/L～6.0mg/L NAA, emergence rate, all higher than 40%, can further screen (seeing Figure 1A).
second takes turns hormone combination screening, and concrete steps are as follows:
(1) at FS, fs, CMS, multitude elm B, assorted 37 first rice crops in Yunnan, gather in latter 7 days, intercept respectively FS, fs, CMS, multitude elm B, the assorted 37 first rice crop acrial parts in Yunnan with falling 1～fall the rice straw of 3 node dormancy buds, clean;
(2) respectively by the rice straw in FS, fs step (1) Suo Shu, CMS, multitude elm B, Yunnan assorted 37, with the hormonal composition of the variable concentrations shown in table 3～table 6, soak 48h processing;
(3) rice straw with node dormancy bud after step (2) processing is evenly spread out over to earthing on seedling bed, by the conventional seedbed of paddy rice method of raising seedling, grows seedlings and manage;
(4) treat that the 3 leaf-4 leaf phases of regrowth add up respectively emergence rate, further screening obtains suitable combination and concentration (in Table 3～table 6 and Fig. 2～Fig. 5) thereof.
Result is found (see figure 2): (0.1mg/L 2 for DK hormonal composition, 4-D+Kinetin), with 2 of 0.1mg/L, 4-D is quantitative, only has the emergence rate of Kinetin concentration CMS when 2.5mg/L～3.5mg/L higher than 40%, remaining emergence rate is all lower than 40%, and result is all undesirable, so consider, casts out DK hormonal composition.
Result is found (see figure 3): NB hormonal composition (0.1mg/L NAA+6-BA), take the NAA of 0.1mg/L for quantitative, and when 6-BA concentration is during at 10.0mg/L～12.0mg/L, the emergence rate of 5 kinds of materials is all between 40%～56%; When 6-BA concentration is during lower than 10.0mg/L, only has the emergence rate of CMS higher than 40%.Considering 0.1mg/L NAA+10.0mg/L～12.0mg/L 6-BA induction emergence rate better, all higher than 40%, is the suitable for plant hormone combinations proportioning of screening acquisition.
Result is found (see figure 4): BN hormonal composition (0.25mg/L 6-BA+NAA), take the 6-BA of 0.25mg/L for quantitative, and when NAA concentration is during at 30.0mg/L～60.0mg/L, the emergence rate of 5 kinds of materials is all between 40%～67%; When NAA concentration is during higher than 60.0mg/L, only have CMS NAA concentration be 60.0mg/L～120 mg/L and Yunnan assorted 37 emergence rate that is 60.0mg/L～90mg/L in NAA concentration higher than 40%, all the other concentration and material emergence rate are all lower than 40%.Considering 0.25mg/L 6-BA+30.0mg/L～60.0mg/L NAA induction emergence rate better, all higher than 40%, is the suitable for plant hormone combinations proportioning of screening acquisition.
BN hormonal composition is according to the combination of new proportioning after the first round screening relatively good single plant hormone NAA of acquisition and 6-BA, why do not continue the single hormone of screening is to consider to use single hormone to induce the growth efficiency of new root and sleeping bud limited simultaneously, and use therefore combines them.
Result is found (see figure 5): KN hormonal composition (10mg/L Kinetin+ NAA), take the Kinetin of 10mg/L for quantitative, and when NAA concentration is during at 2.0 mg/L～5.0mg/L, the emergence rate of 5 kinds of materials is between 40%～65%; When NAA concentration is during lower than 2.0mg/L, the emergence rate in fs, CMS and Yunnan assorted 37 is higher than 40%, and the emergence rate of FS and multitude elm B is lower than 40%; When NAA concentration is greater than 5.0mg/L, the emergence rate in fs, FS, CMS and Yunnan assorted 37 is higher than 40%, and the emergence rate of multitude elm B is lower than 40%.Considering 10mg/L Kinetin+ 2.0 mg/L～5.0mg/L NAA induction emergence rate better, all higher than 40%, is the suitable for plant hormone combinations proportioning of screening acquisition.
The assorted 37 5 kinds of materials in comprehensive fs, FS, CMS, multitude elm B and Yunnan are considered:
0.1mg/L NAA+10.0 mg/L～12.0mg/L 6-BA, 0.25mg/L 6-BA+30 mg/L～60mg/L NAA or 10mg/L Kinetin+ 2.0 mg/L～5.0mg/L NAA emergence rate are higher, for suitable plant hormone combination matching, all can induce with the rice straw of sleeping bud and emerge, obtain the good effect of emergence rate between 40%～67%.
embodiment 3
Select syzygy 42B(conventional Rice maintainer), multitude elm B(conventional Rice maintainer), the assorted 38(hybrid F in Yunnan
1) first rice crop acrial part with falling 1～rice straw of 3 node dormancy buds of falling is test material, by hormone solution 0.25mg/L 6-BA+30mg/L NAA immersion treatment 48 hours, and to take the seed sprouting seedling in syzygy 42B, multitude elm B, Yunnan assorted 38 be contrast.3 leaf-4 leaf phases were transplanted to experimental field, by the conventional field production management method of paddy rice, carried out field production management.Investigate and compare emergence rate, the dry matter of the Sheng of tillering phase, leaf area index and chlorophyll content, spike length when ripe, Number of kernels etc., investigate the superiority of hormone induction regeneration.
Concrete grammar is as described below:
1) in syzygy 42B, multitude elm B, Yunnan, assorted 37 first rice crops gather ins in latter 7 days, intercepting syzygy 42B, multitude elm B, the assorted 38 first rice crop acrial parts in Yunnan with falling 1～fall the rice straw of 3 node dormancy buds, clean;
2) rice straw described in step 1) is used respectively solution 0.25mg/L 6-BA+30mg/L NAA immersion treatment 48 hours;
3) rice straw with node dormancy bud after processing is evenly spread out over to earthing on seedling bed, sows under the same conditions syzygy 42B, multitude elm B, assorted 38 seeds in Yunnan simultaneously for contrasting, all by the conventional seedbed of paddy rice method of raising seedling, cultivate and manage;
4) the 3 leaf-4 leaf phases of rice shoot for the treatment of regrowth and seed sprouting are transplanted to experimental field, and add up emergence rate;
5) by the conventional field production management method of paddy rice, carry out field production management after turning green, investigate dry matter, leaf area index and the chlorophyll content of the Sheng phase of tillering, when ripe, investigate spike length, Number of kernels.
Result of the test shows (table 7): adopt conventional Rice maintainer and hybrid rice with falling 1～fall the rice straw of 3 node dormancy buds in conjunction with hormone solution immersion treatment, although regrowth emergence rate is lower than directly using the emergence rate of seed germination, but every Related Agronomic Characters in each material later stage: dry matter, plant height, spike length, Number of kernels, and leaf area index, the materials variances of chlorophyll content and seed sprouting is not remarkable, show that the present invention adopts band to fall 1～fall the conventional Rice maintainer of 3 node dormancy buds and the rice straw of hybrid rice and all can keep suitable Main Agronomic Characters with conventional seed growing in conjunction with the method for hormone solution-treated, can be normally solid, but the inventive method does not need to carry out hybrid seeding repeatedly, not only avoided the expensive of the paddy rice cross breeding production of hybrid seeds, shorten breeding cycle, can also well keep its hybrid vigour.
embodiment 4
With fs, CMS, multitude elm B, the assorted 37(hybrid F in Yunnan
1) first rice crop acrial part with falling 1～rice straw of 3 node dormancy buds of falling is test material, use respectively hormonal composition immersion treatment, and be contrast with the rice stub in fs, CMS, multitude elm B, Yunnan assorted 37, in process of the test, investigate relatively emergence rate, the plant height of the Sheng of tillering phase, dry matter, leaf area index and chlorophyll content, spike length when ripe, grain husk is spent number etc., investigates the superiority of hormone induction regeneration.
Concrete grammar is as described below:
1) at fs, CMS, multitude elm B, the assorted 37(hybrid F in Yunnan
1) first rice crop gathered in latter 7 days, intercepts respectively fs, CMS, multitude elm B, the assorted 37(hybrid F in Yunnan
1) first rice crop acrial part with falling 1～fall the rice straw of 3 node dormancy buds, clean;
2) rice straw described in step 1) is used respectively solution 0.25mg/L 6-BA+30mg/L NAA immersion treatment 48 hours;
3) by after processing with falling 1～rice straw of 3 node dormancy buds of falling is evenly spread out over earthing on seedling bed, by the conventional seedbed of paddy rice method of raising seedling, grows seedlings and manage;
4) treat regrowth 3 leaf～4 leaf phases transplanting land for growing field crops, under identical field condition, transplanting fs, CMS, multitude elm B, Yunnan assorted 37 leaves down 1～falls the rice stub of 3 node dormancy buds for contrasting, process and contrast all and carry out field production management by the conventional field production management method of paddy rice, and add up emergence rate.
Investigate that test is processed and to impinging upon plant height, dry matter, the leaf area index of the Sheng phase of tillering, when ripe, investigate spike length, grain husk is spent number.
Result of the test shows (table 8): utilize emergence rate, leaf area index, the plant height of the regeneration of hormone induction rice straw Nodes sleeping bud to be all extremely significantly better than the rice stub that ratoon rice is directly transplanted; Spike length be significantly longer than contrast and show significantly sparse, so grain husk spend number be extremely significantly better than contrast.
Embodiment 4 shows: for special rice material (as female sterile, male sterile) and conventional Rice, hybrid rice (hybrid F
1) rice material get with falling 1～fall 3 node dormancy buds rice straw in conjunction with HORMONE TREATMENT after emergence rate than the emergence rate of the contrast of direct transplanting rice pile material, improved 112.3%～132.9%, dry matter, leaf area index, plant height, spike length, grain husk spend number to increase substantially than contrast, difference is extremely remarkable, has produced unforeseeable technique effect.
Comprehensive embodiment 1～4 result draws, for special rice material (educate as female, female sterile, male sterile etc.) and conventional Rice, hybrid rice (hybrid F
1) adopt with falling 1～fall 3 node dormancy buds rice straw by HORMONE TREATMENT or need not hormone-treated regeneration efficiency be all extremely significantly better than simulating ratoon rice and directly transplant rice stub regeneration, its emergence rate, leaf area index, plant height, grain husk are spent and are counted, the contrast of Number of kernels geometric ratio increases substantially, difference is extremely remarkable, solve special material and be difficult for a difficult problem of breeding, do not need to carry out hybrid seeding repeatedly, expensive, the shortening breeding cycle of the paddy rice cross breeding production of hybrid seeds have not only been avoided, can also well keep its hybrid vigour, produce unforeseeable technique effect.
Claims (2)
1. paddy rice sleeping bud is bred the method for regeneration plant, comprises the following steps:
(1) at first rice crop, gathered in latter 7 days, intercepting paddy rice acrial part with falling 1～fall the rice straw of 3 node dormancy buds;
(2) rice straw step (1) Suo Shu is cleaned to the rear plant hormone composition of using and soaked 40～50 hours, then be spread out on seedling bed and earthing, by the conventional seedbed of paddy rice method of raising seedling, grow seedlings and manage; The formula of described plant hormone composition is 0.25mg/L 6-BA+30～60mg/L NAA, 0.1mg/L NAA+10.0～12.0mg/L 6-BA or 10.0mg/L Kinetin+2.0～5.0mg/L NAA;
(3) treat regrowth 3 leaf～4 leaf phases transplanting land for growing field crops, by the conventional field production management method of paddy rice, carry out field production management.
2. paddy rice sleeping bud according to claim 1 is bred the method for regeneration plant, it is characterized in that: the formula of described plant hormone composition is 0.25mg/L 6-BA+30mg/L NAA.
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| CN111374007A (en) * | 2020-04-15 | 2020-07-07 | 黑龙江省农业科学院绥化分院 | Breeding F for increasing tillering regeneration capacity of rice1Method for generation harvest number |
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