CN103743915A - Identification method for high-quality seedling of rubber tree - Google Patents
Identification method for high-quality seedling of rubber tree Download PDFInfo
- Publication number
- CN103743915A CN103743915A CN201410024749.XA CN201410024749A CN103743915A CN 103743915 A CN103743915 A CN 103743915A CN 201410024749 A CN201410024749 A CN 201410024749A CN 103743915 A CN103743915 A CN 103743915A
- Authority
- CN
- China
- Prior art keywords
- bud
- rubber tree
- endogenous hormones
- stereotype
- juvenile form
- 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.)
- Granted
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/415—Assays involving biological materials from specific organisms or of a specific nature from plants
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Biotechnology (AREA)
- Endocrinology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Pretreatment Of Seeds And Plants (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention relates to an identification method for a high-quality seedling and particularly relates to an identification method for a high-quality seedling of a rubber tree. The identification method comprises the following steps: respectively measuring the contents of auxin, gibberellin, mitogen and abscisic acid with old buds and young buds as materials, wherein the old buds and the young buds are located according to the previous experience; analyzing the contents of the auxin, the gibberellin, the mitogen and the abscisic acid and proportional difference to obtain numerical values of endogenous hormones with significant differences of the old buds and the young buds; measuring and calculating the numerical values of the endogenous hormones with significant differences of buds to be measured, and comparing the numerical values with a threshold value to judge whether the buds to be measured are young buds; and if the buds to be measured are the young buds, identifying the obtained seedling as a high-quality seedling. According to the identification method for the high-quality seedling of the rubber tree, provided by the invention, an identification method for the old buds and the young buds located according to the previous experience is updated to be an endogenous hormone quantification identification technology, so that the identification result is objective and effective, the requirements on the background knowledge and actual operation of people are low, and the identification can be favorably implemented.
Description
Technical field
The present invention relates to the authentication method of high quality seedling, in particular to a kind of authentication method of rubber tree high quality seedling.
Background technology
The main mode that adopts budling of production plantation of rubber tree, budling is mainly comprised of stock and scion two parts.Stock is generally chosen the kind of better resistance, and the kind that output is high, product are of fine quality is generally chosen in scion.Bud point in scion, according to the difference of stage of development, is divided into stereotype bud and juvenile form bud.Through forefathers' investigation and analysis, juvenile form bud bud grafting tree is better than stereotype bud bud grafting tree, and the seedling that juvenile form bud bud grafting tree obtains shows the proterties of fast growing and high yield.
The early stage research of stereotype bud and the juvenile form bud criteria for classifying mainly concentrates on morphological index, as being purple, trunk, juvenile form bud is the standard that the stage of development changes that is considered to such as conical, the plant with these features or one Partial Feature is considered to juvenile form, and what do not have these features is just considered to stereotype.According to this morphological criteria, China and the scholar of Ceylon adopt and are less than 3-4 Ya Hedi position, the seedling tree high-order bit bud in age, have carried out the contrast experiment of a series of juvenile form, stereotype planting material output, and they believe that juvenile form material can compare high yield.But experimental result has negated unexpectedly their supposition: the output of juvenile form and stereotype material does not show obvious difference.
Chinese scholar Liu in 1958 sees that the morphological criteria taking the lead in to the rubber tree stage of development changes has proposed suspection, and points out: the upper and lower (bud) of 2 years raw seedlings may be all in juvenile form stage (Liu Naijian.Rubber breeding.The chief editor of South China College of Tropical Crops.Tropical crops thremmatology, Beijing: agriculture publishing house, 1991,213-222).In segmentation rubber tapping experiment afterwards, they find that 7-8 raw rubber tree trunk Chan Jiao capacity variation district is at 2.5-3m, and the variation zone of 40 years raw trees is at 4.5-6m.Liu Song spring is being summed up on the basis of domestic and international experimental data subsequently, when the age criterion that rubber tree has been proposed to divide the stage of development is seedling tree 1-4, the upper and lower position of plant (bud) all belongs to juvenile form, in the time of 6 years, tree crown part (bud) belongs to stereotype, but the following position of trunk 1m (bud) still keeps juvenile form.Because rubber tree generally blooms at 5-6, in fact the sexal maturity standard of this standard and development of plants phasic change is basically identical, this result has also obtained checking in peroxidase enzymes and Esterase Isozyme determination experiment, and current most of scholar holds approval attitude to this standard.
But, forefathers have just done the experience location at age and position to rubber tree juvenile form bud and stereotype bud, locate by rule of thumb juvenile form bud and stereotype bud and learn that to be whether the high quality seedling of rubber tree require height to the background knowledge of appraiser and practical operation, be unfavorable for the carrying out of identifying.
Summary of the invention
The object of the present invention is to provide a kind of authentication method of rubber tree high quality seedling, to solve the above problems.
The authentication method of a kind of rubber tree high quality seedling providing in an embodiment of the present invention, comprises the following steps:
(a) a plurality of juvenile form buds that, the rubber tree of the same kind of the many strains of random acquisition is determined according to existing experience and the bud point of a plurality of stereotype buds;
(b) measure, respectively the mean value of the content of every kind of endogenous hormones in the bud point of described juvenile form bud and described stereotype bud and every kind of endogenous hormones of every kind of bud point, and the content of every kind of endogenous hormones is done to difference analysis;
(c), according to the mean value of the proportionate relationship value between the endogenous hormones of the proportionate relationship value between the endogenous hormones of every kind of bud point of the every strain rubber tree of cubage of every kind of endogenous hormones in step (b) and every kind of bud point, and the proportionate relationship value between the endogenous hormones of every kind of bud point is done to difference analysis;
(d), according to the difference analysis result of step (b) and step (c), determine endogenous hormones kind and the proportionate relationship with significant difference;
(e), to thering is endogenous hormones kind and the proportionate relationship of significant difference, the mean value that step (b) and step (c) are obtained calculates respectively, obtains the mean value of the mean value of two kinds of bud points, and as boundary value;
(f), measure and calculate endogenous hormones and the corresponding numerical value of proportionate relationship that bud to be measured has significant difference, and institute's value and boundary value are compared, judge whether it is juvenile form bud, if belong to juvenile form bud, the seedling obtaining is accredited as high quality seedling.
Preferably, in described step (e), to thering is endogenous hormones kind and the proportionate relationship of significant difference, in the mean value that step (b) and step (c) are obtained, 2-4 numerical value of the percentage difference maximum of two kinds of bud points calculates, obtain the mean value of the mean value of two kinds of bud points, and as boundary value.
Preferably, the endogenous hormones kind of stating in step (b) and step (c) comprises abscisic acid, auxin, mitogen and gibberellin.
Preferably, described juvenile form bud is the juvenile form bud before bud grafting, and described stereotype bud is the stereotype bud before bud grafting; Described bud to be measured is taken from the bud before bud grafting.
Preferably, the bud of described juvenile form bud point is taken from the stabilization vane branch of the rubber tree that was less than for 6 ages.
Preferably, the bud of described stereotype bud point is taken from the tree crown of the rubber tree that was greater than for 6 ages.
Preferably, described tree crown be in under the stabilization vane branch of several the 3rd fluffy leaves up.
Preferably, described rubber tree is that rubber tree kind GT1, cloud grind 77-2 and Yun Yan 77-4.
Preferably, described juvenile form bud is after the bud grafting of juvenile form bud survives, 7-10 days after 3-5cm saw rod above healing place of bud grafting point, the bud of healing place;
Described stereotype bud is after the bud grafting of stereotype bud survives, 7-10 days after 3-5cm saw rod above healing place of bud grafting point, the bud of healing place;
Described bud to be measured is taken from after bud grafting survives, and bud grafting is put to 7-10 days after the 3-5cm saw rod of healing place top, the bud of healing place.
Preferably, described juvenile form bud and described stereotype bud are taken from respectively rubber tree kind GT1, cloud grinds 77-2 and Yun Yan 77-4, and bud grafting is to rubber tree kind GT1.
Endogenous hormones is that plant intensive amount is small, the active substance of adjustable plant physiology reaction, they are at aspects such as cell division and elongation, tissue and Organ Differentiation, Florescence and seed set, ripe and old and feeble, dormancy and sprouting and in vitro tissue cultivations, respectively or mutually the growth of regulating plant, growth and break up in phase.What budling was focused on is the quickly-healing of callus after bud grafting, so the endogenous hormones relevant with it is auxin, gibberellin, mitogen and abscisic acid substantially, auxin promotes the Growth and Differentiation of cell and the formation of callus; Gibberellin can accelerate cell division, promotes to sprout; Mitogen plays activation in promoting cell division; Abscisic acid promotes dormancy, suppresses the Proliferation and differentiation of cell.According to the coordinative role principle of endogenous hormones, the growth conditions of bud can not only be inferred from the content of single hormone, and need to analyze the ratio between hormone, therefore, the content of abscisic acid, auxin, mitogen and gibberellin and mutual ratio have been carried out to difference analysis, obtaining auxin, gibberellin, mitogen and abscisic acid has a great difference in content and the ratio of stereotype bud and juvenile form bud, thereby can be used as the fast-growing of Budding In Hevea shoot survival percent, budling and the standard that bud grafting tree produces glue ability.
It is material that stereotype bud and the juvenile form bud of forefathers' experience location take in the present invention, measure respectively auxin, gibberellin, mitogen and ABA content, by analyzing the difference analysis of auxin, gibberellin, mitogen and ABA content and mutual ratio thereof, stereotype bud and the juvenile form bud authentication method of forefathers' experience location are risen to endogenous hormones quantification authenticate technology, qualification result is more objective effectively, low to personnel's background knowledge and practical operation requirement, be beneficial to the enforcement of evaluation.
Embodiment
Below by specific embodiment, the present invention is described in further detail.
Embodiment 1:
The authentication method of a kind of rubber tree high quality seedling providing in an embodiment of the present invention, comprises the following steps:
(a) a plurality of juvenile form buds that, the rubber tree of the same kind of the many strains of random acquisition is determined according to existing experience and the bud point of a plurality of stereotype buds;
(b) measure, respectively the mean value of the content of every kind of endogenous hormones in the bud point of described juvenile form bud and described stereotype bud and every kind of endogenous hormones of every kind of bud point, and the content of every kind of endogenous hormones is done to difference analysis;
(c), according to the mean value of the proportionate relationship value between the endogenous hormones of the proportionate relationship value between the endogenous hormones of every kind of bud point of the every strain rubber tree of cubage of every kind of endogenous hormones in step (b) and every kind of bud point, and the proportionate relationship value between the endogenous hormones of every kind of bud point is done to difference analysis;
(d), according to the difference analysis result of step (b) and step (c), determine endogenous hormones kind and the proportionate relationship with significant difference;
(e), to thering is endogenous hormones kind and the proportionate relationship of significant difference, the mean value that step (b) and step (c) are obtained calculates respectively, obtains the mean value of the mean value of two kinds of bud points, and as boundary value;
(f), measure and calculate endogenous hormones and the corresponding numerical value of proportionate relationship that bud to be measured has significant difference, and institute's value and boundary value are compared, judge whether it is juvenile form bud, if belong to juvenile form bud, the seedling obtaining is accredited as high quality seedling.
Endogenous hormones is that plant intensive amount is small, the active substance of adjustable plant physiology reaction, they are at aspects such as cell division and elongation, tissue and Organ Differentiation, Florescence and seed set, ripe and old and feeble, dormancy and sprouting and in vitro tissue cultivations, respectively or mutually the growth of regulating plant, growth and break up in phase.What budling was focused on is the quickly-healing of callus after bud grafting, so the endogenous hormones relevant with it is auxin, gibberellin, mitogen and abscisic acid substantially, auxin promotes the Growth and Differentiation of cell and the formation of callus; Gibberellin can accelerate cell division, promotes to sprout; Mitogen plays activation in promoting cell division; Abscisic acid promotes dormancy, suppresses the Proliferation and differentiation of cell.According to the coordinative role principle of endogenous hormones, the growth conditions of bud can not only be inferred from the content of single hormone, and need to analyze the ratio between hormone, therefore, the content of abscisic acid, auxin, mitogen and gibberellin and mutual ratio have been carried out to difference analysis, obtaining auxin, gibberellin, mitogen and abscisic acid has a great difference in content and the ratio of stereotype bud and juvenile form bud, thereby can be used as the boundary value of fast-growing and the bud grafting tree product glue ability of Budding In Hevea shoot survival percent, budling.
It is material that stereotype bud and the juvenile form bud of forefathers' experience location take in the present invention, measure respectively auxin, gibberellin, mitogen and ABA content, by analyzing the difference analysis of auxin, gibberellin, mitogen and ABA content and mutual ratio thereof, stereotype bud and the juvenile form bud authentication method of forefathers' experience location are risen to endogenous hormones quantification authenticate technology, and qualification result is more objective effectively.
The physiological activity of stabilization vane branch is more stable, representative.
The rubber tree of the same kind of many strains is that the data in order to obtain have statistical significance; Similarly, a plurality of juvenile form buds and a plurality of stereotype bud are also that data in order to obtain have statistical significance.The content of the endogenous hormones in the rubber tree of different cultivars and between proportional difference different.
Preferably, described juvenile form bud is the juvenile form bud before bud grafting, and described stereotype bud is the stereotype bud before bud grafting; Described bud to be measured is taken from the bud before bud grafting.
Preferably, described juvenile form bud is after the bud grafting of juvenile form bud survives, 7-10 days after 3-5cm saw rod above healing place of bud grafting point, the bud of healing place;
Described stereotype bud is after the bud grafting of stereotype bud survives, 7-10 days after 3-5cm saw rod above healing place of bud grafting point, the bud of healing place;
Described bud to be measured is taken from after bud grafting survives, and bud grafting is put to 7-10 days after the 3-5cm saw rod of healing place top, the bud of healing place.
Bud be divided into the bud of directly directly getting from stabilization vane branch before bud grafting and bud bud grafting is survived the bud got; Correspondingly, the boundary value that the bud of directly directly getting from stabilization vane branch before bud grafting if adopt obtains, identifies that when whether bud to be measured is high quality seedling, bud to be measured directly obtains from branch; The boundary value that the bud of getting after bud bud grafting being survived if adopt obtains, identifies when whether bud to be measured is high quality seedling, after bud bud grafting to be measured survives, obtains this bud.
Preferably, the bud of described juvenile form bud point is taken from the stabilization vane branch of the rubber tree that was less than for 6 ages.
Preferably, the bud of described stereotype bud point is taken from the tree crown of the rubber tree that was greater than for 6 ages.
Preferably, described tree crown be in under the stabilization vane branch of several the 3rd fluffy leaves up.
Become age tree to be generally 6 years above trees, empirical evidence, is greater than bud in the tree crown of rubber tree at 6 ages in stereotype.
The breeding material that rubber tree kind GT1 is rubber tree, cloud grinds 77-2 and Yun Yan 77-4 is the conventional high gumwood in Yunnan Province.Therefore, these 3 rubber tree kinds are representative.Preferably, rubber tree is that rubber tree kind GT1, cloud grind 77-2 and Yun Yan 77-4.
Particularly,
The collection of juvenile form bud: in rubber tree propagation garden, rubber tree kind GT1, cloud are ground to each kind of 77-2 and Yun Yan 77-4 and choose at random 100 strains, get scion in stabilization vane branch, port wax sealing, takes back laboratory.
The collection of stereotype bud: each kind that 6 ages, above rubber tree kind GT1, the cloud that becomes tree in age ground 77-2 and Yun Yan 77-4 is chosen to 100 strains at random, in under up the stabilization vane branch of several the 3rd fluffy leaves get scion, port wax sealing, takes back laboratory.
Be sample time after every day, the sun rose, on rubber tree leaf, not having after dew is sampling, is probably between point-10 in the mornings 9; Can not sample rainy day.
The juvenile form bud of fetching and stereotype bud are got to band bud point bud sheet, the bud length of a film 0.3cm obtaining, wide 0.3cm with pocket knife.10 bud sheets are 1 sample, grind the mensuration of carrying out endogenous hormones auxin, gibberellin, mitogen and abscisic acid.Every strain rubber tree is got 30 bud sheets, and every strain repeats for 3 times.
The extraction of bud endogenous hormones:
1, reagent
Extract: accurately take 0.22g2,6-di-tert-butyl-4-methy phenol (BHT), is dissolved in 800ml methyl alcohol, and adding distil water is settled to 1000ml;
Dilution, accurately takes sodium chloride (NaCl) 8g, ten dihydrated sodium hydrogen phosphate (Na
2hPO
312H
2o) 2.96g, is dissolved in 1000ml distilled water, then adds 1ml tween and 1g gelatin, and low-grade fever is dissolved.
2, extraction step:
A, sample is put to mortar, added about 0.3g silica sand, be ground to powdered form, the gradation of 2ml extract is added to mortar, all proceed in 2.5ml centrifuge tube;
B, by centrifuge tube in sky 4 ℃, beautiful CT15RT hydro-extractor, centrifugal 15 minutes of 4000rmp, get supernatant;
C, supernatant is put into C18 extraction column filter, then put into traditional vacuum concentrate drying device (LNG-T96) concentrated, to as the oil drop sample of small Semen setariae grain size, add dilution to be settled to 1ml, obtain extract.
3, the mensuration of endogenous hormones:
Extract obtained above is carried out respectively to the mensuration of the content of abscisic acid, auxin, mitogen and gibberellin.The mensuration of abscisic acid, auxin, mitogen and GA content adopts respectively enzyme linked immunosorbent assay, and the ELISA kit of abscisic acid, auxin, mitogen and gibberellin is provided by Shanghai Yuan Ye biotechnology company.The mensuration of abscisic acid, auxin, mitogen and gibberellin operates according to the instructions of corresponding ELISA kit.
It is as shown in table 1 that rubber tree kind GT1, cloud grind the content of the juvenile form bud of 77-2 and Yun Yan 77-4 and the abscisic acid (ABA) in stereotype bud, auxin (IAA), mitogen (CTK) and gibberellin (GA), and standard deviation is the standard deviation between the sample of 100 strains.
Table 1 rubber tree is old, juvenile form bud bud point endogenous hormones table
To the proportionate relationship value between the content value of every kind of endogenous hormones of 100 strain samples and the endogenous hormones of every kind of bud point, adopt DPS software to carry out difference analysis, obtain table 2 and table 3.
Table 2 rubber tree is old, juvenile form bud bud point endogenous hormones analysis of variance table
P<0.05 is significant difference, and p<0.01 is utmost point significant difference.
As can be seen from Table 2, hormone-content aspect is analyzed, and between old juvenile form bud, except GA, IAA, CTK, ABA difference all reach the utmost point level of signifiance; Between kind, except IAA, GA, CTK, ABA difference all reach the level of signifiance.
Table 3 rubber tree is old, juvenile form bud bud point endogenous hormones ratio analysis of variance table
Table 3 can be found out, the analysis of endogenous hormones ratio aspect, and between old juvenile form bud, except IAA/CTK, between all the other endogenous hormones, ratio IAA/GA, IAA/ABA, GA/CTK, GA/ABA, CTK/ABA all reach significant difference level; Between kind, each endogenous hormones proportional difference all reaches the level of signifiance.
By the result of table 2 and table 3 difference analysis, obtain the kind of the endogenous hormones with the significance difference opposite sex except GA and IAA/CTK, simultaneously the numerical value difference between the average of juvenile form bud and the average of stereotype bud in associative list 1.Numerical value difference adopts the ratio between large numerical value and little numerical value to represent, ratio is larger, represents that difference is larger; The ratio of numerical value is not considered standard deviation, obtains numerical value as shown in table 4.
Table 4 rubber tree is old, juvenile form bud endogenous hormones ratio table
| Hormone kind | GT1 ratio | Cloud grinds 77-2 ratio | Cloud grinds 77-4 ratio |
| IAA | 1.2453 | 1.82698 | 1.5745 |
| GA | 1.2603 | 1.0308 | 1.0152 |
| CTK | 1.4813 | 1.6856 | 1.1576 |
| ABA | 1.8766 | 1.3177 | 3.0181 |
| IAA/GA | 1.01206 | 1.7724 | 1.5510 |
| IAA/CTK | 1.1846 | 1.0842 | 1.3524 |
| IAA/ABA | 2.3030 | 2.4182 | 4.7568 |
| GA/CTK | 1.1778 | 1.6301 | 1.1395 |
| GA/ABA | 2.3556 | 1.3551 | 3.0577 |
| CTK/ABA | 2.7797 | 2.2210 | 3.4937 |
As known from Table 4, for rubber tree kind GT1, choose the average of juvenile form bud corresponding to CTK/ABA, GA/ABA, IAA/ABA and ABA difference and stereotype bud as boundary value, obtaining rubber tree kind GT1 is high quality seedling: CTK/ABA is greater than 0.8004, GA/ABA is greater than 0.000755, IAA/ABA is greater than 0.00054, ABA and is less than 124.8000ng/ml;
Similarly, for rubber tree kind cloud, grind 77-2, choose the average of juvenile form bud corresponding to IAA/ABA, CTK/ABA and IAA difference and stereotype bud as boundary value, obtaining rubber tree kind cloud, to grind 77-2 be high quality seedling: IAA/ABA is greater than 0.00188, CTK/ABA is greater than 1.86309, IAA and is greater than 0.0735ng/ml;
Similarly, for rubber tree kind cloud, grind 77-4, choose the average of juvenile form bud corresponding to IAA/ABA, CTK/ABA, GA/ABA and ABA difference and stereotype bud as boundary value, obtaining rubber tree kind cloud, to grind 77-2 be high quality seedling: IAA/ABA is greater than 0.001065, CTK/ABA is greater than 0.79601, GA/ABA is greater than 0.00106, ABA and is less than 84.42ng/ml.
Get at random 50 juvenile form buds and 50 the stereotype buds of rubber tree kind GT1, measure respectively the content of abscisic acid, auxin, mitogen and gibberellin, carry out data analysis, all meet the restriction of boundary value.
Same method validation rubber tree kind cloud grinds juvenile form bud and the stereotype bud of 77-2 and Yun Yan 77-4, also all meets the restriction of boundary value.
Embodiment 2:
Take kind GT1 as stock, and to grind 77-4, GT1 be scion with becoming to set age above kind 77-2, cloud 6 ages in rubber tree propagation garden; After bud grafting survives, 7-10 days after 3-5cm saw rod above healing place of bud grafting point, gets bud grafting healing place bud sheet, the bud length of a film 0.3cm obtaining, and wide 0.3cm, 10 is 1 sample, every strain repeats for 3 times.Similarly, respectively each rubber tree kind of stereotype bud and juvenile form bud is chosen to 100 strains at random.
Be sample time after every day, the sun rose, on rubber tree leaf, not having after dew is sampling, is probably between point-10 in the mornings 9; Can not sample rainy day.
Within after saw rod 7-10 days, are the most vigorous periods of callus physiological activity, callus is the most vigorous position of physiological activity after budling saw rod.Now, can judge according to the content of endogenous hormones the upgrowth situation of the bud after bud grafting.
With the method in embodiment 1, obtain bud sheet and it is carried out to the mensuration of abscisic acid, auxin, mitogen and GA content.
Take from rubber tree kind GT1, cloud and grind the juvenile form bud of 77-2 and Yun Yan 77-4 and the bud grafting of stereotype bud to kind GT1, after bud grafting survives, after saw rod 7-10 days, get bud grafting healing place bud sheet, the content that obtains abscisic acid in bud sheet (ABA), auxin (IAA), mitogen (CTK) and gibberellin (GA) is as shown in table 5, and standard deviation is the standard deviation between the sample of 100 strains.
Table 5 rubber tree is old, the endogenous hormones table of juvenile form budling saw rod healing place bud point
To the proportionate relationship value between the content value of every kind of endogenous hormones of 100 strain samples and the endogenous hormones of every kind of bud point, adopt DPS software to carry out difference analysis, obtain table 6 and table 7.
The endogenous hormones analysis of variance table of bud grafting healing place bud point after table 6 Budding In Hevea seedling saw rod
P<0.05 is significant difference, and p<0.01 is utmost point significant difference.
As can be seen from Table 6, Endogenous Hormone Contents in Vitro aspect is analyzed, and between old juvenile form bud, endogenous hormones GA, IAA, CTK, ABA difference all reach the utmost point level of signifiance; Between kind, endogenous IAA, GA, CTK, ABA difference all reach the utmost point level of signifiance.
The endogenous hormones ratio analysis of variance table of bud grafting healing place bud point after table 7 Budding In Hevea seedling saw rod
Table 7 can be found out, the analysis of endogenous hormones ratio aspect, and between old juvenile form bud, except CTK/ABA, between all the other endogenous hormones, ratio IAA/GA, IAA/CTK, IAA/ABA, GA/CTK, GA/ABA all reach significant difference level; Between kind, each endogenous hormones proportional difference all reaches the utmost point level of signifiance.
By the result of table 6 and table 7 difference analysis, obtain having the kind of the related endogenous hormones of significant difference, simultaneously the numerical value difference between the average of juvenile form bud and the average of stereotype bud in associative list 5.Numerical value difference adopts the ratio between large numerical value and little numerical value to represent, ratio is larger, represents that difference is larger; The ratio of numerical value is not considered standard deviation, obtains numerical value as shown in table 8.
The endogenous hormones ratio table of bud grafting healing place bud point after table 8 Budding In Hevea seedling saw rod
| Hormone kind | GT1 ratio | Cloud grinds 77-2 ratio | Cloud grinds 77-4 ratio |
| IAA | 1.0303 | 1.0325 | 1.1547 |
| GA | 1.6578 | 1.6393 | 1.2635 |
| CTK | 1.2816 | 1.2493 | 1.1538 |
| ABA | 1.0580 | 1.0782 | 1.0873 |
| IAA/GA | 1.6185 | 1.4601 | 1.7026 |
| IAA/CTK | 1.1818 | 1.2778 | 1.2727 |
| IAA/ABA | 1.1000 | 1.2600 | 1.1471 |
| GA/CTK | 1.3030 | 1.0933 | 1.3208 |
| GA/ABA | 1.7517 | 1.1634 | 1.5203 |
| CTK/ABA | 1.3564 | 1.0613 | 1.1586 |
As known from Table 8, for obtaining scion to rubber tree kind GT1 from rubber tree kind GT1, after bud grafting survives, 7-10 days after saw rod, get the bud sheet of bud grafting healing place, choose the average of juvenile form bud corresponding to GA/ABA, GA and IAA/GA difference and stereotype bud as boundary value, the bud that meets following condition is high quality seedling: GA/ABA is greater than 0.00205, GA is greater than 0.06525ng/ml, and IAA/GA is less than 0.163435;
Similarly, for obtaining scion to rubber tree kind GT1 from rubber tree kind 77-2, after bud grafting survives, 7-10 days after saw rod, get the bud sheet of bud grafting healing place, choose the average of juvenile form bud corresponding to GA and IAA/GA difference and stereotype bud as boundary value, what meet following condition is greater than 0.0644ng/ml for high quality seedling: GA, and IAA/GA is less than 0.263675;
Similarly, for obtaining scion to rubber tree kind GT1 from rubber tree kind 77-4, after bud grafting survives, 7-10 days after saw rod, get the bud sheet of bud grafting healing place, choose the average of juvenile form bud corresponding to IAA/GA and GA/ABA difference and stereotype bud as boundary value, what meet following condition is less than 0.20705 for high quality seedling: IAA/GA, and GA/ABA is greater than 0.001865.
Get at random 50 juvenile form buds and 50 the stereotype buds of rubber tree kind GT1, bud grafting is to rubber tree kind GT1 respectively, after bud grafting survives, 7-10 days after saw rod, get the bud sheet of bud grafting healing place, the content of measuring abscisic acid, auxin, mitogen and gibberellin, carries out data analysis, all meets the restriction of boundary value.
Get at random juvenile form bud and stereotype bud that rubber tree kind cloud grinds 77-2 and Yun Yan 77-4, and difference bud grafting is to rubber tree kind GT1, after bud grafting survives, 7-10 days after saw rod, get respectively the bud sheet of bud grafting healing place, measure respectively the content of abscisic acid, auxin, mitogen and gibberellin, carry out data analysis, also all meet the restriction of boundary value.
In sum, the method for evaluation juvenile form bud provided by the invention and stereotype bud, the evaluation that is quantification by experience positioning and lifting, method is more objective effectively, low to personnel's background knowledge and practical operation requirement, is beneficial to the enforcement of evaluation.
The foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, for a person skilled in the art, the present invention can have various modifications and variations.Within the spirit and principles in the present invention all, any modification of doing, be equal to replacement, improvement etc., within all should being included in protection scope of the present invention.
Claims (10)
1. an authentication method for rubber tree high quality seedling, is characterized in that, comprises the following steps:
(a) a plurality of juvenile form buds that, the rubber tree of the same kind of the many strains of random acquisition is determined according to existing experience and the bud point of a plurality of stereotype buds;
(b) measure, respectively the mean value of the content of every kind of endogenous hormones in the bud point of described juvenile form bud and described stereotype bud and every kind of endogenous hormones of every kind of bud point, and the content of every kind of endogenous hormones is done to difference analysis;
(c), according to the mean value of the proportionate relationship value between the endogenous hormones of the proportionate relationship value between the endogenous hormones of every kind of bud point of the every strain rubber tree of cubage of every kind of endogenous hormones in step (b) and every kind of bud point, and the proportionate relationship value between the endogenous hormones of every kind of bud point is done to difference analysis;
(d), according to the difference analysis result of step (b) and step (c), determine endogenous hormones kind and the proportionate relationship with significant difference;
(e), to thering is endogenous hormones kind and the proportionate relationship of significant difference, the mean value that step (b) and step (c) are obtained calculates respectively, obtains the mean value of the mean value of two kinds of bud points, and as boundary value;
(f), measure and calculate endogenous hormones and the corresponding numerical value of proportionate relationship that bud to be measured has significant difference, and institute's value and boundary value are compared, judge whether it is juvenile form bud, if belong to juvenile form bud, the seedling obtaining is accredited as high quality seedling.
2. the authentication method of a kind of rubber tree high quality seedling according to claim 1, it is characterized in that, in described step (e), to thering is endogenous hormones kind and the proportionate relationship of significant difference, in the mean value that step (b) and step (c) are obtained, 2-4 numerical value of the percentage difference maximum of two kinds of bud points calculates, obtain the mean value of the mean value of two kinds of bud points, and as boundary value.
3. the authentication method of a kind of rubber tree high quality seedling according to claim 1, is characterized in that, the endogenous hormones kind in described step (b) and step (c) comprises abscisic acid, auxin, mitogen and gibberellin.
4. the authentication method of a kind of rubber tree high quality seedling according to claim 1, is characterized in that, described juvenile form bud is the juvenile form bud before bud grafting, and described stereotype bud is the stereotype bud before bud grafting; Described bud to be measured is taken from the bud before bud grafting.
5. the authentication method of a kind of rubber tree high quality seedling according to claim 4, is characterized in that, the bud point of described juvenile form bud is taken from the stabilization vane branch of the rubber tree that was less than for 6 ages.
6. the authentication method of a kind of rubber tree high quality seedling according to claim 5, is characterized in that, the bud point of described stereotype bud is taken from the tree crown of the rubber tree that was greater than for 6 ages.
7. the authentication method of a kind of rubber tree high quality seedling according to claim 6, is characterized in that, described tree crown be in under the stabilization vane branch of several the 3rd fluffy leaves up.
8. the authentication method of a kind of rubber tree high quality seedling according to claim 7, is characterized in that, the kind of rubber tree is that GT1, cloud grind 77-2 and Yun Yan 77-4.
9. the authentication method of a kind of rubber tree high quality seedling according to claim 1, is characterized in that, described juvenile form bud is after the bud grafting of juvenile form bud survives, 7-10 days after 3-5cm saw rod above healing place of bud grafting point, the bud of healing place;
Described stereotype bud is after the bud grafting of stereotype bud survives, 7-10 days after 3-5cm saw rod above healing place of bud grafting point, the bud of healing place;
Described bud to be measured is taken from after bud grafting survives, and bud grafting is put to 7-10 days after the 3-5cm saw rod of healing place top, the bud of healing place.
10. the authentication method of a kind of rubber tree high quality seedling of stating according to claim 9, is characterized in that, described juvenile form bud and described stereotype bud are taken from respectively rubber tree kind GT1, cloud grinds 77-2 and Yun Yan 77-4, and difference bud grafting is to rubber tree kind GT1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410024749.XA CN103743915B (en) | 2014-01-20 | 2014-01-20 | The authentication method of rubber tree high quality seedling |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410024749.XA CN103743915B (en) | 2014-01-20 | 2014-01-20 | The authentication method of rubber tree high quality seedling |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103743915A true CN103743915A (en) | 2014-04-23 |
| CN103743915B CN103743915B (en) | 2015-09-30 |
Family
ID=50500950
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410024749.XA Active CN103743915B (en) | 2014-01-20 | 2014-01-20 | The authentication method of rubber tree high quality seedling |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103743915B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107389594A (en) * | 2017-06-30 | 2017-11-24 | 北京农业信息技术研究中心 | A kind of stem stalk terminal bud recognition positioning method and device based on terahertz light |
| CN108254510A (en) * | 2018-03-12 | 2018-07-06 | 云南省热带作物科学研究所 | A kind of method that quick discriminating rubber tree juvenile form bud connects clone nursery stock |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101120641A (en) * | 2006-08-11 | 2008-02-13 | 中国热带农业科学院热带生物技术研究所 | Seedling budding method for rubber tree minitype bud sticks |
| WO2012099100A1 (en) * | 2011-01-18 | 2012-07-26 | 住友ゴム工業株式会社 | Method for producing isoprenoid, isoprenoid, callus, method for inducing callus, and method for culturing callus |
-
2014
- 2014-01-20 CN CN201410024749.XA patent/CN103743915B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101120641A (en) * | 2006-08-11 | 2008-02-13 | 中国热带农业科学院热带生物技术研究所 | Seedling budding method for rubber tree minitype bud sticks |
| WO2012099100A1 (en) * | 2011-01-18 | 2012-07-26 | 住友ゴム工業株式会社 | Method for producing isoprenoid, isoprenoid, callus, method for inducing callus, and method for culturing callus |
Non-Patent Citations (2)
| Title |
|---|
| 张治礼等: "树木发育阶段理论及其在橡胶树育种中的应用", 《(热带亚热带植物学报》, vol. 9, no. 3, 30 March 2001 (2001-03-30), pages 262 - 268 * |
| 李明谦等: "橡胶树不同类型芽片的嫁接繁殖试验", 《热带农业科技》, vol. 34, no. 1, 31 January 2001 (2001-01-31), pages 4 - 6 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107389594A (en) * | 2017-06-30 | 2017-11-24 | 北京农业信息技术研究中心 | A kind of stem stalk terminal bud recognition positioning method and device based on terahertz light |
| CN108254510A (en) * | 2018-03-12 | 2018-07-06 | 云南省热带作物科学研究所 | A kind of method that quick discriminating rubber tree juvenile form bud connects clone nursery stock |
| CN108254510B (en) * | 2018-03-12 | 2020-05-01 | 云南省热带作物科学研究所 | Method for rapidly identifying young bud-grafting clonal seedlings of rubber trees |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103743915B (en) | 2015-09-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Niu et al. | The compensation effects of physiology and yield in cotton after drought stress | |
| Du et al. | Altitudinal patterns of leaf stoichiometry and nutrient resorption in Quercus variabilis in the Baotianman Mountains, China | |
| Wang et al. | Physiological response of soybean plants to water deficit | |
| Suriyagoda et al. | Growth and phosphorus nutrition of rice when inorganic fertiliser application is partly replaced by straw under varying moisture availability in sandy and clay soils | |
| Zhang et al. | Effects of arbuscular mycorrhizal fungi on the drought tolerance of Cyclobalanopsis glauca seedlings under greenhouse conditions | |
| Lopez Del Egido et al. | A spectrophotometric assay for robust viability testing of seed batches using 2, 3, 5-triphenyl tetrazolium chloride: using Hordeum vulgare L. as a model | |
| Adjel et al. | Salt stress effects on seed germination and seedling growth of barley (Hordeum Vulgare L.) genotypes | |
| CN102577798A (en) | Method for identifying salt tolerance of corn | |
| Daneshvar et al. | Stimulation of germination in dormant seeds of Juniperus polycarpos by stratification and hormone treatments | |
| Liu et al. | Influence of pericarp, cotyledon and inhibitory substances on sharp tooth oak (Quercus aliena var. acuteserrata) germination | |
| Suga et al. | Pathogenic characters of Japanese potato strains of Ralstonia solanacearum | |
| Gonçalves et al. | Assessment of growth and yield performance of rubber tree clones of the IAC 500 series | |
| Chen et al. | Effects of arbuscular mycorrhizal inoculation on the growth, zinc distribution and photosynthesis of two citrus cultivars grown in low-zinc soil | |
| Lv et al. | Comparative response to drought in primitive and modern wheat: a cue on domestication | |
| Yan et al. | Effects of drought stress on water content and biomass distribution in summer maize (Zea mays L.) | |
| Najafi Vafa et al. | Soil microorganisms and seaweed application with supplementary irrigation improved physiological traits and yield of two dryland wheat cultivars | |
| CN103650948A (en) | Identification method for soybean drought resistance | |
| Liu et al. | Evaluation of salinity tolerance and genetic diversity of thirty-three switchgrass (Panicum virgatum) populations | |
| CN103743915B (en) | The authentication method of rubber tree high quality seedling | |
| Nascimento-Júnior et al. | Indirect selection of maize genotypes based on associations between root agronomic and anatomical characters | |
| Sailaja et al. | Validation of Oryza2000 model under combined nitrogen and water limited situations | |
| Gyuleva et al. | Five years growth of Paulownia on two sites in Bulgaria. | |
| CN108037248B (en) | Identification method of selenium-rich millet | |
| Ran et al. | Effects of indole-3-butytric acid on lead and zinc accumulations in Pseudostellaria maximowicziana | |
| JAMES et al. | Rates of production and internal levels of ethylene in the vegetative cotton plant |
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 |