CN103743915B - The authentication method of rubber tree high quality seedling - Google Patents
The authentication method of rubber tree high quality seedling Download PDFInfo
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- CN103743915B CN103743915B CN201410024749.XA CN201410024749A CN103743915B CN 103743915 B CN103743915 B CN 103743915B CN 201410024749 A CN201410024749 A CN 201410024749A CN 103743915 B CN103743915 B CN 103743915B
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- 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
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- 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
Abstract
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.The stereotype bud that the present invention locates with forefathers' experience and juvenile form bud are for material, measure auxin, gibberellin, mitogen and ABA content respectively, by analyzing the difference analysis of auxin, gibberellin, mitogen and ABA content and mutual ratio thereof, obtain the numerical value that stereotype bud and juvenile form bud have the endogenous hormones of significant difference, measure and calculate the endogenous hormones numerical value that bud to be measured has significant difference, and itself and boundary value are compared, judge it whether as juvenile form bud, if belong to juvenile form bud, the seedling obtained then is accredited as high quality seedling.The stereotype bud that forefathers' experience is located by the authentication method of rubber tree high quality seedling provided by the invention and juvenile form bud authentication method rise to endogenous hormones quantification authenticate technology, qualification result is more effectively objective, require low to the background knowledge of personnel and practical operation, be beneficial to the enforcement of qualification.
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 adopting budling of production plantation of rubber tree, budling is mainly made up of stock and scion two parts.Stock generally chooses 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 the investigation and analysis of forefathers, juvenile form bud bud grafting tree is better than stereotype bud bud grafting tree, and the proterties that the seedling obtained shows fast growing and high yield is set in the bud grafting of juvenile form bud.
The early stage research of stereotype bud and the juvenile form bud criteria for classifying mainly concentrates on morphological index, as juvenile form bud be purple, trunk conically etc. is considered to the standard that the stage of development changes, the plant namely 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 Ceylon scholar adopt Ya Hedi position, the seedling tree high-order bit bud being less than 3-4 age, and carried out the contrast experiment of a series of juvenile form, stereotype planting material output, 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 taken the lead in the rubber tree stage of development changes proposes suspection, and points out: the upper and lower (bud) of 2 years raw seedlings may all be in juvenile form stage (Liu Naijian.Rubber breeding.South China College of Tropical Crops edits.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 on the basis of summing up domestic and international experimental data subsequently, when the age criterion and seedling tree 1-4 that divide the stage of development are proposed to rubber tree, the upper and lower position of plant (bud) all belongs to juvenile form, when 6 years, tree crown part (bud) belongs to stereotype, but trunk 1m still keeps juvenile form with lower portion (bud).Because rubber tree generally blooms at 5-6, in fact the sexal maturity standard that changes of this standard and plant developing stage is basically identical, this result also obtains 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 juvenile form bud and stereotype bud by rule of thumb 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 provided in an embodiment of the present invention, comprises the following steps:
The bud point of a multiple juvenile form bud that the rubber tree of the same kind of the many strains of (), random acquisition is determined according to existing experience and multiple stereotype bud;
(b), measure described juvenile form bud and described stereotype bud respectively bud point in the mean value of the content of often kind of endogenous hormones and often kind of endogenous hormones of often kind of bud point, and difference analysis is done to the content of often kind of endogenous hormones;
(c), mean value according to the proportionate relationship value between the endogenous hormones of the proportionate relationship value between the endogenous hormones of often kind of bud point of the every strain rubber tree of the cubage of often kind of endogenous hormones in step (b) and often kind of bud point, and difference analysis is done to the proportionate relationship value between the endogenous hormones of often kind of bud point;
D (), difference analysis result according to step (b) and step (c), determine endogenous hormones kind and the proportionate relationship with significant difference;
(e), to endogenous hormones kind and the proportionate relationship with significant difference, mean value step (b) and step (c) 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 bud to be measured there is the endogenous hormones of significant difference and the numerical value corresponding to proportionate relationship, and institute's value and boundary value are compared, judge it whether as juvenile form bud, if belong to juvenile form bud, the seedling obtained then is accredited as high quality seedling.
Preferably, in described step (e), to endogenous hormones kind and the proportionate relationship with significant difference, 2-4 the numerical value that in the mean value obtain step (b) and step (c), the percentage difference of two kinds of bud points is maximum calculates, obtain the mean value of the mean value of two kinds of bud points, and as boundary value.
Preferably, the endogenous hormones kind stated 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 takes from the bud before bud grafting.
Preferably, the bud point of described juvenile form bud takes from the stabilization vane branch of the rubber tree being less than for 6 ages.
Preferably, the bud point of described stereotype bud takes from the tree crown of the rubber tree being greater than for 6 ages.
Preferably, described tree crown be under the stabilization vane branch of up several 3rd fluffy leaf.
Preferably, described rubber tree is rubber tree kind GT1, cloud grinds 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, by 7-10 days after 3-5cm saw rod above healing place of bud grafting point, and the bud of healing place.
Preferably, described juvenile form bud and described stereotype bud take from rubber tree kind GT1 respectively, 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 in cell division and elongation, to organize and in Organ Differentiation, Florescence and seed set, ripe and old and feeble, Dormancy and bourgeon and excised cotyledon etc., respectively or mutually in phase regulating plant growth, grow and break up.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, and the Growth and Differentiation of auxin promotion cell and calli induction; Gibberellin can accelerate cell division, promotes to sprout; Mitogen plays activation in promotion cell division; Abscisic acid then promotes dormancy, the Proliferation and differentiation of T suppression cell.According to the coordinative role principle of endogenous hormones, the growth conditions of bud can not only estimate from the content of single hormone, and the ratio between Water demand hormone, therefore, the content of abscisic acid, auxin, mitogen and gibberellin and mutual ratio have been carried out difference analysis, obtain auxin, gibberellin, mitogen and abscisic acid and have a great difference at the content of stereotype bud and juvenile form bud and ratio, thus the standard that Budding In Hevea shoot survival percent, the fast-growing of budling and bud grafting tree produce glue ability can be used as.
The stereotype bud that the present invention locates with forefathers' experience and juvenile form bud are for material, measure auxin, gibberellin, mitogen and ABA content respectively, by analyzing the difference analysis of auxin, gibberellin, mitogen and ABA content and mutual ratio thereof, the stereotype bud of locate forefathers' experience and juvenile form bud authentication method rise to endogenous hormones quantification authenticate technology, qualification result is more effectively objective, require low to the background knowledge of personnel and practical operation, be beneficial to the enforcement of qualification.
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 provided in an embodiment of the present invention, comprises the following steps:
The bud point of a multiple juvenile form bud that the rubber tree of the same kind of the many strains of (), random acquisition is determined according to existing experience and multiple stereotype bud;
(b), measure described juvenile form bud and described stereotype bud respectively bud point in the mean value of the content of often kind of endogenous hormones and often kind of endogenous hormones of often kind of bud point, and difference analysis is done to the content of often kind of endogenous hormones;
(c), mean value according to the proportionate relationship value between the endogenous hormones of the proportionate relationship value between the endogenous hormones of often kind of bud point of the every strain rubber tree of the cubage of often kind of endogenous hormones in step (b) and often kind of bud point, and difference analysis is done to the proportionate relationship value between the endogenous hormones of often kind of bud point;
D (), difference analysis result according to step (b) and step (c), determine endogenous hormones kind and the proportionate relationship with significant difference;
(e), to endogenous hormones kind and the proportionate relationship with significant difference, mean value step (b) and step (c) 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 bud to be measured there is the endogenous hormones of significant difference and the numerical value corresponding to proportionate relationship, and institute's value and boundary value are compared, judge it whether as juvenile form bud, if belong to juvenile form bud, the seedling obtained then is accredited as high quality seedling.
Endogenous hormones is that plant intensive amount is small, the active substance of adjustable plant physiology reaction, they in cell division and elongation, to organize and in Organ Differentiation, Florescence and seed set, ripe and old and feeble, Dormancy and bourgeon and excised cotyledon etc., respectively or mutually in phase regulating plant growth, grow and break up.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, and the Growth and Differentiation of auxin promotion cell and calli induction; Gibberellin can accelerate cell division, promotes to sprout; Mitogen plays activation in promotion cell division; Abscisic acid then promotes dormancy, the Proliferation and differentiation of T suppression cell.According to the coordinative role principle of endogenous hormones, the growth conditions of bud can not only estimate from the content of single hormone, and the ratio between Water demand hormone, therefore, the content of abscisic acid, auxin, mitogen and gibberellin and mutual ratio have been carried out difference analysis, obtain auxin, gibberellin, mitogen and abscisic acid and have a great difference at the content of stereotype bud and juvenile form bud and ratio, thus the boundary value that Budding In Hevea shoot survival percent, the fast-growing of budling and bud grafting tree produce glue ability can be used as.
The stereotype bud that the present invention locates with forefathers' experience and juvenile form bud are for material, measure auxin, gibberellin, mitogen and ABA content respectively, by analyzing the difference analysis of auxin, gibberellin, mitogen and ABA content and mutual ratio thereof, the stereotype bud of locate forefathers' experience and juvenile form bud authentication method rise to endogenous hormones quantification authenticate technology, and qualification result is more effectively objective.
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, multiple juvenile form bud and multiple 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 takes 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, by 7-10 days after 3-5cm saw rod above healing place of bud grafting point, and the bud of healing place.
Bud to be divided into before bud grafting the bud directly directly got from stabilization vane branch and the bud got after bud bud grafting being survived; Correspondingly, according to the boundary value that the bud directly directly got from stabilization vane branch before bud grafting obtains, when identifying whether bud to be measured is high quality seedling, bud to be measured directly obtains from branch; According to the boundary value that the bud got after bud bud grafting being survived obtains, when identifying whether bud to be measured is high quality seedling, after bud bud grafting to be measured survives, obtain this bud.
Preferably, the bud point of described juvenile form bud takes from the stabilization vane branch of the rubber tree being less than for 6 ages.
Preferably, the bud point of described stereotype bud takes from the tree crown of the rubber tree being greater than for 6 ages.
Preferably, described tree crown be under the stabilization vane branch of up several 3rd fluffy leaf.
Become set the tree being generally more than 6 years age, empirical evidence, the bud be greater than in the tree crown of the rubber tree at 6 ages is in stereotype.
Rubber tree kind GT1 is the breeding material of rubber tree, and it is the high gumwood that Yunnan Province is commonly used that cloud grinds 77-2 and Yun Yan 77-4.Therefore, these 3 rubber tree kinds are representative.Preferably, rubber tree is rubber tree kind GT1, cloud grinds 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 random selecting 100 strain of 77-2 and Yun Yan 77-4, get scion in stabilization vane branch, port wax sealing, takes back laboratory.
The collection of stereotype bud: each kind random selecting 100 strain of the rubber tree kind GT1 more than 6 ages becoming set age, cloud being ground to 77-2 and Yun Yan 77-4, under the stabilization vane branch of up several 3rd fluffy leaf get scion, port wax sealing, takes back laboratory.
After being the sun rise every day sample time, being sampling after rubber tree leaf not having dew, is probably between at 9 .-10 in the morning; Rainy day can not sample.
Band bud point bud sheet is got to the juvenile form bud fetched and stereotype bud pocket knife, the bud length of a film 0.3cm obtained, wide 0.3cm.10 bud sheets are 1 sample, and the mensuration of endogenous hormones auxin, gibberellin, mitogen and abscisic acid is carried out in grinding.Every strain rubber tree gets 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), be dissolved in 800ml methyl alcohol, 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 to be put in mortar, add about 0.3g silica sand, be ground to powdered form, the gradation of 2ml extract is added mortar, all proceed in 2.5ml centrifuge tube;
B, by centrifuge tube in sky beautiful 4 DEG C, CT15RT hydro-extractor, centrifugal 15 minutes of 4000rmp, get supernatant;
C, supernatant put into C18 extraction column and filter, then put into traditional vacuum concentrate drying device (LNG-T96) and concentrate, to the oil drop sample as small Semen setariae grain size, add dilution and be settled to 1ml, obtain extract.
3, the mensuration of endogenous hormones:
Extract obtained above is carried out respectively the mensuration of the content of abscisic acid, auxin, mitogen and gibberellin.The mensuration of abscisic acid, auxin, mitogen and GA content adopts enzyme linked immunosorbent assay respectively, 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.
The content that rubber tree kind GT1, cloud grind abscisic acid (ABA), auxin (IAA), mitogen (CTK) and gibberellin (GA) in the juvenile form bud of 77-2 and Yun Yan 77-4 and stereotype bud is as shown in table 1, 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
Adopt DPS software to carry out difference analysis to the proportionate relationship value between the content value of often kind of endogenous hormones of 100 strain samples and the endogenous hormones of often kind of bud point, 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 pole 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 reaches the pole level of signifiance; Between kind except IAA, GA, CTK, ABA difference all reaches 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, endogenous hormones ratio aspect is analyzed, 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 with the endogenous hormones of the significance difference opposite sex except GA and IAA/CTK, the numerical value difference simultaneously in associative list 1 between the average of juvenile form bud and the average of stereotype bud.Numerical value difference adopts the ratio between large numerical value and little numerical value to represent, ratio is larger, then represent that difference is larger; The ratio of numerical value does not consider 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 CTK/ABA, GA/ABA, IAA/ABA and ABA juvenile form bud corresponding respectively 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, 77-2 is ground for rubber tree kind cloud, choose the average of IAA/ABA, CTK/ABA and IAA juvenile form bud corresponding respectively 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, 77-4 is ground for rubber tree kind cloud, choose the average of IAA/ABA, CTK/ABA, GA/ABA and ABA juvenile form bud corresponding respectively 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 50 juvenile form buds and 50 stereotype buds of rubber tree kind GT1 at random, measure the content of abscisic acid, auxin, mitogen and gibberellin respectively, 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:
With kind GT1 for stock, rubber tree propagation garden and more than 6 ages become to set kind 77-2 age, to grind 77-4, GT1 be scion to cloud; After bud grafting survives, above healing place of bud grafting point, 7-10 days after 3-5cm saw rod, gets bud grafting healing place bud sheet, the bud length of a film 0.3cm obtained, wide 0.3cm, and 10 is 1 sample, and every strain repeats for 3 times.Similarly, respectively to each rubber tree kind random selecting 100 strain of stereotype bud and juvenile form bud.
After being the sun rise every day sample time, being sampling after rubber tree leaf not having dew, is probably between at 9 .-10 in the morning; Rainy day can not sample.
Within after saw rod 7-10 days, are callus physiological activity the most vigorous periods, callus is the position that after budling saw rod, physiological activity is the most vigorous.Now, the upgrowth situation of the bud after bud grafting can be judged according to the content of endogenous hormones.
Obtain bud sheet with the method in embodiment 1 and it carried out to the mensuration of abscisic acid, auxin, mitogen and GA content.
Take from rubber tree kind GT1, juvenile form bud that cloud grinds 77-2 and Yun Yan 77-4 and the bud grafting of stereotype bud be to kind GT1, after bud grafting survives, after saw rod 7-10 days, get bud grafting healing place bud sheet, the content obtaining abscisic acid (ABA) in bud sheet, 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
Adopt DPS software to carry out difference analysis to the proportionate relationship value between the content value of often kind of endogenous hormones of 100 strain samples and the endogenous hormones of often kind of bud point, 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 pole 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 reaches the pole level of signifiance; Between kind, endogenous IAA, GA, CTK, ABA difference all reach the pole 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, endogenous hormones ratio aspect is analyzed, 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 pole level of signifiance.
By the result of table 6 and table 7 difference analysis, obtain the kind of the endogenous hormones had involved by significant difference, the numerical value difference simultaneously in associative list 5 between the average of juvenile form bud and the average of stereotype bud.Numerical value difference adopts the ratio between large numerical value and little numerical value to represent, ratio is larger, then represent that difference is larger; The ratio of numerical value does not consider 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, scion is obtained to rubber tree kind GT1 for 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 GA/ABA, GA and IAA/GA juvenile form bud corresponding respectively and stereotype bud as boundary value, the bud meeting 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, scion is obtained to rubber tree kind GT1 for 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 GA and IAA/GA juvenile form bud corresponding respectively 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, scion is obtained to rubber tree kind GT1 for 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, the average choosing IAA/GA and GA/ABA juvenile form bud corresponding respectively and stereotype bud is as boundary value, and what meet following condition is greater than 0.001865 for high quality seedling: IAA/GA is less than 0.20705, GA/ABA.
Get 50 juvenile form buds and 50 stereotype buds of rubber tree kind GT1 at random, 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, measure the content of abscisic acid, auxin, mitogen and gibberellin, carry out data analysis, all meet the restriction of boundary value.
Get juvenile form bud and stereotype bud that rubber tree kind cloud grinds 77-2 and Yun Yan 77-4 at random, and difference bud grafting is to rubber tree kind GT1, after bud grafting survives, 7-10 days after saw rod, get the bud sheet of bud grafting healing place respectively, measure the content of abscisic acid, auxin, mitogen and gibberellin respectively, carry out data analysis, also all meet the restriction of boundary value.
In sum, the method for qualification juvenile form bud provided by the invention and stereotype bud, experience location is risen to the qualification of quantification, and method is more effectively objective, requires low, be beneficial to the enforcement of qualification to the background knowledge of personnel and practical operation.
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 amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (9)
1. an authentication method for rubber tree high quality seedling, is characterized in that, comprises the following steps:
The bud point of a multiple juvenile form bud that the rubber tree of the same kind of the many strains of (), random acquisition is determined according to existing experience and multiple stereotype bud;
(b), measure described juvenile form bud and described stereotype bud respectively bud point in the mean value of the content of often kind of endogenous hormones and often kind of endogenous hormones of often kind of bud point, and difference analysis is done to the content of often kind of endogenous hormones;
(c), mean value according to the proportionate relationship value between the endogenous hormones of the proportionate relationship value between the endogenous hormones of often kind of bud point of the every strain rubber tree of the cubage of often kind of endogenous hormones in step (b) and often kind of bud point, and difference analysis is done to the proportionate relationship value between the endogenous hormones of often kind of bud point;
D (), difference analysis result according to step (b) and step (c), determine endogenous hormones kind and the proportionate relationship with significant difference;
(e), to endogenous hormones kind and the proportionate relationship with significant difference, mean value step (b) and step (c) 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 bud to be measured there is the endogenous hormones of significant difference and the numerical value corresponding to proportionate relationship, and institute's value and boundary value are compared, judge it whether as juvenile form bud, if belong to juvenile form bud, the seedling obtained then is accredited as high quality seedling;
Described endogenous hormones is abscisic acid, auxin, mitogen and gibberellin.
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 endogenous hormones kind and the proportionate relationship with significant difference, 2-4 the numerical value that in the mean value obtain step (b) and step (c), the percentage difference of two kinds of bud points is maximum 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, 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 takes from the bud before bud grafting.
4. the authentication method of a kind of rubber tree high quality seedling according to claim 3, is characterized in that, the bud point of described juvenile form bud takes from the stabilization vane branch of the rubber tree being less than for 6 ages.
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 stereotype bud takes from the tree crown of the rubber tree being greater 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, described tree crown be under the stabilization vane branch of up several 3rd fluffy leaf.
7. the authentication method of a kind of rubber tree high quality seedling according to claim 6, is characterized in that, the kind of rubber tree is GT1, cloud grinds 77-2 and Yun Yan 77-4.
8. 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, by 7-10 days after 3-5cm saw rod above healing place of bud grafting point, and the bud of healing place.
9. the authentication method of a kind of rubber tree high quality seedling stated according to Claim 8, is characterized in that, described juvenile form bud and described stereotype bud take from rubber tree kind GT1 respectively, cloud grinds 77-2 and Yun Yan 77-4, and difference bud grafting is to rubber tree kind GT1.
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| 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 |
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