CN114097445A - Mutation method for peach-apricot intergeneric grafting hybrid - Google Patents
Mutation method for peach-apricot intergeneric grafting hybrid Download PDFInfo
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Abstract
The invention discloses a method for mutagenesis of peach-apricot intergeneric grafting hybrid, which comprises the following steps: selecting apricot plants or peach plants as rootstocks; selecting annual or current-year branches as scions; before grafting, weeding and watering the rootstock, and pretreating the scion; grafting the pretreated scion onto a stock to obtain a grafted seedling; keeping the water content of the soil to be 50-60 wt% after grafting, performing stock shearing and bud picking treatment after the grafting wound is healed, and supplementing exogenous nutrients after the grafting bud grows out; character variation of the grafted seedling is induced by controlling the biomass ratio of the rootstock and the scion; fixing the character of the branch with excellent character variation by sowing, grafting or tissue culture; and (5) expanding the grafted hybrid. The invention can overcome the defects of low hybrid yield, difficult regulation and control of progeny character, long breeding period and the like caused by reproductive isolation barrier in the current distant sexual hybridization, and effectively improves the distant hybridization efficiency between peach genus and apricot genus.
Description
Technical Field
The invention relates to the technical field of peach-apricot intergeneric grafting. In particular to a method for mutagenesis of peach-apricot intergeneric grafting hybrid.
Background
Peach and apricot are the most important fruit and dried fruit trees in the world, both belonging to the Rosaceae (Rosaceae) pruneideae (pruneae Focke), but are different genera, peach being the peach genus (Persica Mill) and apricot being classified in the apricot genus (Armeniaca k. Currently, as for breeding of peach and apricot, seedling selection breeding, cross breeding and molecular assisted breeding are the most important breeding methods at present. The breeding methods have very limited bottleneck for breaking through the unique economic traits of the genus, such as fruit size, self-fruitfulness, fruit color, seed kernel size, kernel size and the like, particularly the quantitative traits. With the improvement of living standard of people, the demand for super fine breed is increasingly outstanding. Therefore, the method for innovating the fruit tree breeding method and quickly cultivating a batch of new fruit tree varieties meeting the human needs have important significance.
The existing research shows that the plant grafting of interspecies (specifices) and the genetic relationship among the interspecies can generate the heritable character variation type expected by breeders, and becomes a new way of germplasm innovation. More and more researches prove that grafting is not only used as a main technical means for fine seedling propagation, germ plasm resource preservation and propagation, but also can cause the characters of grafting chimera, such as leaf color, leaf type, leaf villi, crown height, flowering phase and juvenile phase, stress resistance, yield and quality, change of physiological metabolic process and the like, and the change can be stably transmitted to filial generations. Therefore, grafting becomes a new way of germplasm innovation.
The existing research shows that the cause of the phenomenon may be that the grafting hybridization (Graft Hybrid) is caused by the genetic material exchange between the rootstock and the scion to form a new character, or the adaptive variation is generated by the selective pressure of the grafting hybridization, or the genome and the mitochondrial DNA of both the rootstock and the scion are methylated by the grafting to cause the silencing of related functional genes, and the like.
In recent years, more and more evidence studies have shown that this phenomenon may also be associated with chloroplast, mitochondrial gene lateral transfer, and bidirectional communication of signal molecules such as proteins across the graft site. With the development of research technology, a series of breakthrough research achievements are obtained by exploring the ancient grafting technology, so that the technology developed by the grafting technology based on experience is rapidly developing towards scientific and accurate human controllable characters. The grafting hybridization becomes a new choice in the aspects of innovative germplasm (especially plant breeding of the variety level and above classification status), rich variety, human food safety and the like.
In fruit tree breeding, the creation of super improved varieties through distant hybridization is still a method which is focused on at present and in the future for a long time, however, due to the reproductive isolation barrier among peach and apricot genera, the fruitfulness rate of distant sexual hybridization of peach and apricot is low abnormally, and the bottleneck problems of long breeding period, serious character separation of filial generation, difficult character regulation and control and the like exist. Aiming at the problems, the method breaks through the technical bottleneck of distant hybridization of peach and apricot, improves hybridization affinity, shortens breeding period, and cultivates super improved variety, which is the main way of meeting different requirements of people, reducing production cost and realizing quality and efficiency improvement of industry. However, the mechanism of acquiring genetic traits through interspecific grafting hybridization is complex, and the heritability traits acquired through the plant grafting of the genetic relationship between species and above species still have unpredictability, so that the problems of low hybrid yield, difficult regulation and control of progeny traits, long breeding period and the like of peach-apricot hybridization are solved through the interspecific grafting hybridization mode.
Disclosure of Invention
Therefore, the technical problem to be solved by the invention is to provide a peach-apricot intergeneric grafting hybrid mutagenesis method capable of effectively breaking reproductive isolation barrier between peach and apricot species, so as to solve the bottleneck problems of abnormal low fruit setting rate, long breeding period, serious character separation of hybrid progeny, difficult character regulation and the like of the current peach-apricot distant sexual hybridization.
In order to solve the technical problems, the invention provides the following technical scheme:
a method for mutagenesis of peach-apricot intergeneric grafting hybrid comprises the following steps:
selecting and preparing rootstocks: selecting apricot plants or peach plants as rootstocks;
selecting scions in the step (2): selecting annual or current-year branches as scions; the scion is a peach plant or apricot plant and is of a different species from the stock;
step (3), preparation before grafting: before grafting, weeding and watering the rootstock, and pretreating the scion;
grafting: grafting the pretreated scion onto a stock to obtain a grafted seedling;
step (5), management after grafting: keeping the water content of the soil of the grafted seedling at 50-60 wt% after grafting, performing stock shearing and bud picking treatment after the grafting wound is healed, and supplementing exogenous nutrients after the grafted bud grows out;
step (6) inducing variation: controlling the biomass ratio of the rootstock and the scion by a manual pruning mode to induce the character variation of the grafted seedling to obtain a grafted hybrid;
and (7) fixing characters: fixing the character of the branch with excellent character variation by sowing, grafting or tissue culture;
and (8) expanding the grafted hybrid.
In the method for mutagenesis of peach-apricot intergeneric grafting hybrid, in the step (1), the apricot plant is common apricot (a.vulgaris), siberian apricot (a.sibirica), Liaoxing apricot (a.mandshurica), Tibetan apricot (a.holosericea), purple apricot (a.dyscarppa), Shidan apricot (a.zhidanensis), plum (a.mune), plum apricot (a.limeixing), political apricot (a.zhengheinsis) or Huaren apricot (a.cathayana d.l.fu, b.r.li et j.h.li, sp.nov.);
the Prunus plant is Prunus (Amygdalus L.) or Prunus Amygdalus (subg. Persica L.); the subgenus persicae is fructus Caricae (A.davidiana), Sinkiang fructus Persicae (A.ferganensis), Gansu fructus Persicae (A.kansuensis), semen Juglandis Mandshuricae (A.mira) or fructus Persicae (A.persica); amygdalus belongs to amygdalus, and combinations thereof, and amygdalus. The selection principle of rootstock and scion grafting mutagenesis is as follows: the rootstock with higher utilization rate of nitrogen and the scion with lower utilization rate of nitrogen are used; this is because grafting hybrids require the rootstocks to have higher utilization rates of soil fertilizers and soil moisture, i.e., form a robust root system to support normal production of biomass of the rootstocks and improve environmental suitability.
According to the method for inducing the grafted hybrid between peach and apricot genera, in the step (1), peach trees or apricot trees which are healthy in growth, free of diseases and insect pests and free of gummosis are selected as the stocks, the row spacing of the peach trees or the apricot trees is (2-3) mx (4-8) m [ the stocks are guaranteed to be good in ventilation and light transmission, and meanwhile, the water and fertilizer conditions of the selected stocks in the areas are guaranteed to be high ];
if grafting in spring: selecting a trunk or a backbone branch with the rootstock thickness of more than or equal to 1cm, cutting the trunk or the backbone branch at a position 0.4-1.5 m away from the ground, and cutting the section into a smooth plane for later use;
if grafting in summer: in spring, the trunk or the bone trunk branches with the thickness of more than or equal to 1cm are cut, the current-year new shoots with the thickness of more than 0.6cm are selected as the grafting-planning objects of the scions, and other branches which do not meet the standard are removed.
In the step (1) and the step (2), the rootstocks and the scions used for grafting comprise fruits, seed cores and seed kernels of plants such as closely related species and intergeneric hybrid species of apricot and peach, and tissue and organ life bodies which have independent root systems and have the capability of independently developing into complete plants, namely tissue and organ seedlings.
According to the peach-apricot intergeneric grafting hybrid mutagenesis method, in the step (2), annual or current annual branches without lateral branches which are located at the periphery of a crown and have the thickness of 0.5-1.0 cm are selected, unqualified parts of the tips of the branches are removed and then used as scions (the branches selected as the scions are not limited to the periphery of the crown), and the branches at other parts cannot enter a fruiting period quickly due to the physiological age of bud eyes, so that the effects of high yield and early fruiting are reduced;
in the step (3), the rootstock is watered thoroughly 5-7 days before grafting (the watering time before grafting is not limited to 5-7 days before grafting, the purpose of watering is to keep the water content of soil between 50% and 60% during grafting, promote the flow of sap, and reduce the effect if the water content is not in the range);
in the step (4), grafting is carried out in a mode of grafting by adopting a branch grafting mode or a wood bud grafting mode in spring grafting, and grafting is carried out in a mode of square bud grafting or a wood bud grafting mode in summer grafting; the spring grafting time is from 2 months 15 days to 3 months 20 days after the spring division after the rain, and the summer grafting time is from 6 months 5 days to 6 months 25 days after the summer of the miscanthus. Grafting period includes but is not limited to spring and summer, and grafting in other seasons is easy to cause freezing injury and death in winter due to the fact that the growing period is short and the lignification degree cannot be reached; during grafting, besides three modes of grafting, wood bud grafting and square bud grafting, other methods which can combine the rootstock and the scion, such as cleft grafting, inverted grafting and the like can be selected.
According to the method for inducing the grafting hybrid between peach and apricot genera, the grafting in spring is carried out in a branch grafting mode: cutting the scion into short branches containing 2-3 buds and 5-7 cm in length, cutting the lower end of the scion into a V shape by using a grafting knife, simultaneously cutting a smooth plane of a grafting part of the stock into a corresponding V-shaped notch, or vertically cutting a wound with the depth of 2-3 cm downwards on one side of the grafting part of the stock, and then embedding the scion into the stock; the stock and the scion are tightly bound and tightened by a plastic film to prevent water dispersion and wind break;
grafting in summer by adopting a square bud grafting mode: selecting current-year branches with the thickness of 0.6-1.0 cm from the rootstocks as grafting objects of the scions, and removing barks on the rootstocks and reaching xylem deeply by using parallel double knives with the distance of 0.8-1.0 cm to form rectangular gaps; the buds and bud eyes are obtained from the scion by the same method, the buds and the bud eyes are completely attached to the rectangular gap on the stock and are tightly bound and tightened by a plastic film to prevent water from losing and enable the grafted buds to leak and expose out of the plastic film.
The method for mutagenesis of peach-apricot intergeneric grafting hybrid comprises the following steps of (3): immersing 2-3 cm of a scion base part into the scion pretreatment liquid, and immersing for 30-60 min at the temperature of 20-35 ℃; the scion pretreatment solution is a melatonin solution, or a glucanase solution, or a mixed solution of the melatonin solution and the glucanase solution, and the main purpose of pretreating the scion is to reduce rejection reaction between the stock and the scion and increase grafting affinity; the concentration of the melatonin solution is 10-20 mu mol/L, and the active concentration of the dextranase solution is 50-100U/L.
In the peach-apricot intergeneric grafting hybrid mutagenesis method, in the step (5), after grafting for 3 days, anvil folding is carried out at a position 3-5 cm away from a grafting bud; cutting the stock at the position 1-2 cm above the grafting opening and removing the binding for grafting 7-10 days after summer grafting or 25-30 days after spring graftingAn agent; erasing buds on the rootstock except for the grafted buds, [ promoting growth of the grafted buds ]; when the length of the grafted bud is more than or equal to 10cm, the exogenous nutrients are supplemented for 1-2 times, and the organic matter content of the soil is kept more than or equal to 1.0 wt% [ 3-5 m organic fertilizer can be applied to each mu of land in dormancy season3Keeping the organic matter content of the soil at 1.0 wt% -5.0 wt%; the exogenous nutrient used in the invention is a solid fertilizer (such as a nitrogen fertilizer, a phosphate fertilizer, a potassium fertilizer or a compound fertilizer sold in the market, and the like), and in the solid exogenous nutrient fertilizer, the mass ratio of nitrogen to phosphorus to potassium is 8: 3: 6; besides the nitrogen, phosphorus and potassium elements in the proportion, other single element fertilizers, binary fertilizers, ternary or multi-element compound fertilizers or other fertilizer types rich in medium and trace elements can be added into the exogenous nutrients.
In the method for mutagenesis of peach-apricot intergeneric grafting hybrid, in the step (6), the biomass ratio of new branches germinated on the rootstock and scion grafting buds is controlled to be 1: (0.5-4) to induce the character variation of the grafted seedling. The biomass proportion is measured by the number of branches; the specific method for controlling the new branch of stock germination and the scion bud through manual pruning intervention comprises the following steps: selecting branches with branch angles of 30-60 degrees in east, south, west and north directions of grafted seedlings, and selecting a retention ratio according to the mutual mutagenesis strength of the stocks and the scions: if the variation character of the scion is dominant, 2-4 scion branches and 1 stock branch are reserved, and the rest branches are cut off from the root by using a branch shear; on the contrary, if the variation character of the rootstock is dominant, 2 rootstock branches and 1 scion branch are reserved; meanwhile, the top advantages of the rootstock branches are controlled in a topping mode in the childhood so as to promote the growth of the scion branches; if the two influence each other, the rootstock and the scion keep the same number of branches.
In the method for mutagenesis of peach-apricot intergeneric grafting hybrid, in the step (7), the method for fixing characters and enhancing variation comprises the following steps: collecting the fruit of the grafted hybrid, removing the pulp, and reserving the seed core for sowing;
or, taking the branch generating variation as a scion, and grafting the scion to the same stock for character fixation;
or, the branch generating the variation is taken as the scion and grafted to the xenogeneic rootstock, and the germination of the xenogeneic rootstock is completely limited;
or, carrying out asexual propagation culture by using the variant-producing branches as explants to obtain complete plants. The character fixation is divided into the prior character fixation and the strengthening character fixation, and the prior character fixation needs to be maintained by adopting a method of tissue culture, cuttage or stock grafting; the strengthening character fixation needs to adopt stocks with character characteristics superior to the current variation character for secondary or multiple grafting induction, for example, apricot is taken as scion, and almond, wild peach, Xinjiang peach and the like are taken as strengthening induction signal units of the stocks; the principle of strengthening character fixation in the fixation of the grafted hybrid character is as follows: the dynamic change characteristic of the epigenetic character of the grafted hybrid is influenced by the characters of the stock, and the agronomic characters or the economic characters of the stock are superior to the characters of the grafted hybrid.
In the method for inducing mutation of the peach-apricot intergeneric grafting hybrid, in the step (8), pollen of peach or apricot plants and the grafting hybrid are collected for artificial pollination after the third year of the grafting hybrid and the characters are stable. After the third year character of the grafted hybrid is stable, the grafted hybrid is taken as the minimum unit of breeding, and pollen of the peach or the apricot is collected for distant sexual hybridization, so that the hybrid yield can be improved by more than 2 times. For example, the grafted hybrid peach is taken as a pollination unit, and after the pollen of apricot is selected for treatment, artificial auxiliary pollination hybridization is carried out; or the grafted hybrid apricot is taken as a pollination unit, and the pollen of the peach is selected for treatment and then is subjected to artificial supplementary pollination and hybridization.
The method is also suitable for grafting hybridization among peach genus, apricot genus and plum genus in Prunoideae family, such as Siberian apricot, common apricot, Liaoxing, Tibetan apricot, Shidan apricot, walnut (Prunus mira (Koehne) Kuet Lu), longpeduncle almond, wild peach and the like, and is also suitable for Rosaceae plants, such as cherry and the like. The grafting scions of peaches are grafted to the stocks of apricots, and only the grafted hybrid of the invention has normally grown for more than 6 years from the reported literature.
The grafting hybrid not only has the character variation, but also can obviously reduce the reproductive isolation barrier rate in the sexual hybridization process and improve the yield of hybrid offspring, so the grafting hybrid includes but is not limited to innovative germplasm for distant sexual hybridization or new hybrid range. The method principle and the technology of the invention are suitable for most fruit trees, such as apples, pears, apricots, peaches, dates, walnuts, almonds and the like.
The technical scheme of the invention achieves the following beneficial technical effects:
under the theoretical guidance of stock-ear interaction, epigenetics and the like, through deep research on the peach-apricot grafting hybrid character variation mechanism, on the basis of repeated practice, the invention finds that the character variation of a grafted hybrid is determined by stock selection, scion selection, grafting period, grafting method, post-grafting management, induced variation method, character fixing method and the like. The method can overcome a series of defects of low hybrid yield, difficult regulation and control of progeny character, long breeding period and the like caused by reproductive isolation barrier in the current distant sexual hybridization, effectively improve the distant hybridization efficiency between the peach genus and the apricot genus, realize that the variation phenotype can be generated in the next year of grafting in the current year, shorten the character identification time by more than 1.5 times compared with the character identification time after flowering and fruiting of the progeny of the traditional sexual hybridization, and realize the artificial regulation and control of the grafted hybrid character. The method principle and the technical means of the invention are suitable for most of the existing fruit trees, such as apples, pears, apricots, peaches, dates, walnuts, almonds and the like, and can obviously improve the affinity of distant sexual hybridization and improve the yield of hybrid fruits by more than 3 times.
The invention provides a new technology which is different from the current traditional breeding method between peach and apricot, and can effectively break the reproductive isolation barrier between peach and apricot through a grafted hybrid mutagenesis method. The invention establishes a set of stable, efficient and character-adjustable peach-apricot intergeneric grafting hybrid mutagenesis method, which can break through the self-fixed property of peach and apricot species and obtain expected new germplasm.
Drawings
FIG. 1 shows the performance of apricot on grafted hybrid formed by peach-apricot grafting hybridization in the embodiment of the invention;
FIG. 2 shows the behavior of peach on grafted hybrid formed by peach-apricot grafting hybridization in the embodiment of the invention;
FIG. 3 shows the growth of grafted buds 20-25 days after grafting peach and apricot in example 2 of the present invention;
FIG. 4 shows the growth of grafted buds 20-25 days after grafting peach and apricot according to comparative example 2 of the present invention;
FIG. 5 shows the growth of the grafted bud of peach-apricot in the second 4 months after grafting;
FIG. 6 shows the growth of grafted buds of peach-apricot of comparative example 3 in the second 4 months after grafting.
Detailed Description
The invention is formed by repeated verification on the basis of a reprogramming molecular mechanism which causes the development process of grafted hybrid fruits and is formed by the frequent significant high expression of miRNA, lnRNA, mRNA, cirRNA and the like in scions and rootstocks (Table 1) in the process of systematically researching grafted hybrids between peach and apricot genera. The invention relates to two grafted hybrids, one is to graft the scion of peach genus to the stock of apricot genus to form the grafted hybrid 'apricot + peach'; one is grafting the scion of apricot to the stock of peach to form the grafted hybrid peach and apricot. The two grafting hybrids relate to positive and negative grafting hybrids, and the purpose is to generate peach or apricot grafting hybrids with different characters according to different stocks. Finally, obtaining expected new improved varieties according to whether the variation of the grafted hybrid characters meets the market demand.
TABLE 1 non-coding RNA significantly expressed during peach-apricot grafting hybridization
The technical solution of the present invention is further illustrated by the following specific examples. The 'purple-leaf' flowering peach used in the examples of the present invention is a variation of peach (a. persica) 'youyi' apricot belonging to a production main cultivar of almond apricot (a. cathayana d.l.fu, b.r.li et j.h.li, sp.nov.).
Example 1
Selecting and preparing rootstocks: selecting 'purple leaf' flowering peach trees which grow healthily and have no diseases and insect pests and no gummosis and have the plant spacing of 3m multiplied by 4m, good ventilation and light transmission, high water and fertilizer conditions and the underground water level of less than 1.5m in a plot as rootstocks; the method is implemented in Huang-Huai river basin, and the trunk of the purple-leaf flowering peach stock is cut off in 0.6m away from the ground in the last 3 th month ten days; erasing the sprouts on the rootstock every 7 days, and only keeping sprouts within 15cm at the top end of the rootstock; reserving 1 branch every 80-90 degrees on the current-year-old bone dry branches with the thickness of more than or equal to 0.6cm on the rootstock at 3 days in 6 months, reserving 4 branches as the grafting-planning objects of the scion, and cutting other branches which do not meet the standard from the base;
selecting scions in the step (2): selecting current-year branches positioned at the periphery of a crown as scions, selecting branches which are good-quality and have current-year kernels, have the thickness of 0.6-0.8 cm, are full in bud eyes, have no diseases and pests and do not have lateral branches after kernel-apricot 'good-one' is used for 6 months and 8 days, and removing unqualified parts at two ends of the branches to be used as the scions;
step (3), preparation before grafting: weeding and watering the rootstock 5-7 days before grafting, and pretreating the scion; in the embodiment, the grafted land is subjected to weeding and watering once in 6 months and 4 days; before grafting, soaking a scion base part 2-3 cm into a scion pretreatment solution, wherein the scion pretreatment solution is a mixed solution consisting of a melatonin solution with the concentration of 10 mu mol/L and a glucanase solution with the enzyme activity concentration of 50U/L according to the volume ratio of 1: 1; soaking at the ambient temperature of 20-35 ℃ for 30-60 min, and removing parts of two ends of the branches which do not meet the specification during grafting for later use;
grafting: grafting the pretreated scion onto a stock to obtain a grafted seedling; in this embodiment, the method of "square budding" is used to perform graft induced variation, which specifically includes: the branches reserved on the purple-leaf flowering peach stocks deeply reach xylem by using parallel double knives with the distance of 0.8-1.0 cm, barks are removed to form rectangular gaps, buds and bud eyes are obtained from the 'you one' scions by the same method, the buds and the bud eyes are completely attached to the rectangular gaps of the stocks, and finally, plastic films are used for binding and compacting in a mode of leaking grafted buds, so that the water loss is prevented. Grafting an apricot bud on the branch at the opposite position of the first grafting by adopting the same method;
step (5), management after grafting: keeping the water content of the soil of the grafted seedling at 50-60 wt% after grafting; folding the stock at the position 4cm away from the grafted bud for 4 branches reserved on the stock 3 days after grafting to promote the grafted bud to germinate; cutting a stock at a position 1-2 cm above the grafting opening 7-10 days after grafting and removing a binding object for grafting; every 7 days later, erasing sprouts outside the grafted sprouts; when the length of the grafted bud is more than or equal to 10cm, supplementing exogenous nutrients for 1-2 times, and keeping the organic matter content of the soil to be more than or equal to 1.0 wt%; the exogenous nutrient used in the present example is a solid fertilizer (obtained by mixing commercially available nitrogen fertilizer, phosphate fertilizer, potassium fertilizer, nitrogen-phosphorus-potassium compound fertilizer, or the like), and in the solid exogenous nutrient fertilizer, the mass ratio of nitrogen to phosphorus to potassium is 8: 3: 6; in other embodiments, the exogenous nutrient may contain, in addition to the above-mentioned nitrogen, phosphorus and potassium elements, other single element fertilizers, binary fertilizers, ternary or multicomponent compound fertilizers, or other fertilizer types rich in medium and trace elements;
step (6) inducing variation: the biomass ratio of the rootstock and the scion is controlled by a manual pruning mode to induce the character variation of the grafted seedling; respectively selecting 1-2 branches with branch angles of 30-60 degrees from new branches germinated in east, south, west and north directions of the grafted bud Youyi and the stock bud purple leaf flowering peach, and shearing other redundant branches by using branch shears to enable the grafted bud and the stock bud to form branches with the same biomass, and carrying out grafted hybrid induction;
and (7) fixing characters: fixing the character of the branch with excellent character variation by sowing, grafting or tissue culture; starting the second year after grafting, enhancing water and fertilizer management, removing pulp and preserving seed kernels of collected grafted hybrid fruits, sowing the grafted hybrid fruits into a phenotype determination or molecular marker-assisted selection breeding program of germplasm resources, selecting variant germplasm resources meeting a breeding target, and removing germplasm resources which do not meet requirements; then, the branch generating variation is used as a scion and grafted to a homologus rootstock (the homologus rootstock in the embodiment is 3-year-old 'purple leaf' flowering peach) for character fixation until a mutant with stable characters is generated;
and (8) expanding the grafted hybrid: starting in the third year after grafting, taking a completely developed apricot flower of 'you one' on a grafted hybrid as a female parent of a pollination unit, selecting 'purple leaf' flowering peach pollen with excellent characters as a male parent for pollination and hybridization, increasing the yield of hybrid fruits by 2.5 times compared with the yield of common hybridization (see table 3), collecting the hybrid fruits in the next year, sowing the hybrid fruits and entering a conventional hybridization progeny determination program.
Comparative example 1
Selecting and preparing rootstocks: selecting 'purple leaf' flowering peach trees which grow healthily and have no diseases and insect pests and no gummosis and have the plant spacing of 3m multiplied by 4m, good ventilation and light transmission, high water and fertilizer conditions and the underground water level of less than 1.5m in a plot as rootstocks; the method is implemented in Huang-Huai river basin, and the trunk of the purple-leaf flowering peach stock is cut off in 0.6m away from the ground in the last 3 th month ten days; erasing the sprouts on the rootstock every 7 days, and only keeping sprouts within 15cm at the top end of the rootstock; reserving 1 branch every 80-90 degrees on the current-year-old bone dry branches with the thickness of more than or equal to 0.6cm on the rootstock at 3 days in 6 months, reserving 4 branches as the grafting-planning objects of the scion, and cutting other branches which do not meet the standard from the base;
selecting scions in the step (2): selecting current-year branches positioned at the periphery of a crown as scions, selecting branches which are good-quality and have current-year kernels, have the thickness of 0.6-0.8 cm, are full in bud eyes, have no diseases and pests and do not have lateral branches after kernel-apricot 'good-one' is used for 6 months and 8 days, and removing unqualified parts at two ends of the branches to be used as the scions;
step (3), preparation before grafting: weeding and watering the rootstock 5-7 days before grafting, and pretreating the scion; in the embodiment, the grafted land is subjected to weeding and watering once in 6 months and 4 days; before grafting, soaking a scion base part 2-3 cm in clear water at the ambient temperature of 20-35 ℃ for 30-60 min, and removing parts of two ends of branches which do not meet the specification during grafting for later use;
grafting: grafting the pretreated scion onto a stock to obtain a grafted seedling; in this embodiment, the method of "square budding" is used to perform graft induced variation, which specifically includes: the method comprises the steps of using parallel double knives with the distance of 0.8-1.0 cm to enable branches reserved on 'purple leaf' flowering peach stocks to reach xylem deeply, removing barks to form rectangular gaps, obtaining buds and bud eyes from 'you one' flowering peach scions by the same method, enabling the buds and the bud eyes to be completely attached to the rectangular gaps of the stocks, and finally using plastic films to be bound tightly and firmly in a mode of leaking grafted buds to prevent water from being lost. Grafting an apricot bud on the branch at the opposite position of the first grafting by adopting the same method;
step (5), management after grafting: keeping the water content of the soil of the grafted seedling at 50-60 wt% after grafting, performing stock shearing and bud picking treatment after the grafting wound is healed, and supplementing exogenous nutrients after the grafted bud grows out; folding the stock at the position 4cm away from the grafted bud for 4 branches reserved on the stock 3 days after grafting to promote the grafted bud to germinate; cutting a stock at a position 1-2 cm above the grafting opening 7-10 days after grafting and removing a binding object for grafting; every 7 days later, erasing sprouts outside the grafted sprouts; when the length of the grafted bud is more than or equal to 10cm, supplementing exogenous nutrients for 1-2 times, and keeping the organic matter content of the soil to be more than or equal to 1.0 wt%; the exogenous nutrients used were the same as in example 1; .
Example 2
Selecting and preparing rootstocks: selecting 'you one' apricots in a plot with the physiological age of 6 years, healthy growth, no plant diseases and insect pests, no gummosis, the plant row spacing of 3m multiplied by 4m, good ventilation and light transmission, higher water and fertilizer conditions and the groundwater level of below 1.5m as rootstocks; the method is implemented in Huang-Huai river basin, 2-4 backbone branches with the thickness of more than 1cm and uniform spatial distribution are selected at a position 1.5m away from the ground of a Youyi apricot stock for short-cutting in 3 months and 5 days, the section is kept smooth and flat, and other branches are completely cut from the base;
selecting scions in the step (2): selecting annual branches as scions, selecting branches of purple-leaf flowering peaches which are annual, have the thickness of 0.6-0.8 cm, full bud eyes, no plant diseases and insect pests and no lateral branches and are positioned at the periphery of a crown, and removing unqualified parts at two ends of the branches to be used as the scions;
step (3), preparation before grafting: weeding and watering the rootstock 5-7 days before grafting, and pretreating the scion; in the embodiment, the selected 'you one' apricot stock land is subjected to weeding and watering once in 3 months and 1 day; before grafting, soaking 2-3 cm of a scion base into a scion pretreatment solution, wherein the scion pretreatment solution is a glucanase solution with the enzyme activity of 100U/L, and soaking for 30-60 min at the ambient temperature of 20-35 ℃;
grafting: grafting the pretreated scion onto a stock to obtain a grafted seedling; the grafting induced variation is carried out by adopting a grafting method, which comprises the following steps: firstly cutting a scion into short branches with the length of 5-7 cm and containing 2-3 buds, cutting the lower ends of the branches into V-shaped branches by using a knife, then cutting the smooth section of a backbone branch to be grafted of a stock into V-shaped notches by using a grafting knife, then embedding the scion into the stock, and binding and compacting the scion by using a plastic film to prevent water from losing; if the thickness of the stock backbone branch is larger than 2cm, a scion can be additionally grafted at the relative position of the first scion, and finally the two scions are fixed by plastic cloth to prevent wind fracture.
Step (5), management after grafting: after grafting, timely preventing and controlling the damage of diseases, pests and weeds, keeping the water content of soil of the grafted seedling at 50-60 wt% after grafting, performing stock shearing and bud picking treatment after the grafting wound is healed, and supplementing exogenous nutrients after the grafted bud grows out; checking once every 7 days after grafting, reserving one stock bud at a position corresponding to the grafted branch, and wiping off other buds; removing sprouts outside the grafted sprouts every 7 days after grafting, and promoting the growth of the sprouts; when the length of the grafted bud is more than or equal to 10cm, the exogenous nutrients are supplemented for 1-2 times, and the organic matter content of the soil is kept more than or equal to 1.0 wt% [ 3-5 m organic fertilizer can be applied to each mu of land in dormancy season3Keeping the organic matter content of the soil at 1.0 wt% -5.0 wt%; the exogenous nutrient used in the present example is a solid fertilizer (obtained by mixing commercially available nitrogen fertilizer, phosphate fertilizer, potassium fertilizer, nitrogen-phosphorus-potassium compound fertilizer, or the like), and in the solid exogenous nutrient fertilizer, the mass ratio of nitrogen to phosphorus to potassium is 8: 3: 6 [ in other embodimentsIn the example, the exogenous nutrient can be added with other single element fertilizers, binary fertilizers, ternary or multi-element compound fertilizers or other fertilizer types rich in medium and trace elements besides the nitrogen, phosphorus and potassium elements in the proportion. Compared with the conventional field management, the management method after grafting obviously improves the biomass accumulation (shown in table 2) of the grafting scion and the occurrence of an incompatible event;
step (6) inducing variation: the biomass ratio of the rootstock and the scion is controlled by a manual pruning mode to induce the character variation of the grafted seedling; respectively selecting 1-2 branches with branch angles of 30-60 degrees from new branches sprouting in east, south, west and north directions of the grafting bud and the stock bud, shearing other redundant branches by using a branch shear, enabling the grafting bud and the stock bud to form branches with the same biomass, carrying out grafting hybrid induction, and then merging into conventional field management;
and (7) fixing characters: starting the second year after grafting, enhancing water and fertilizer management, removing pulp and preserving seed kernels of collected grafted hybrid fruits, sowing the grafted hybrid fruits into a phenotype determination or molecular marker-assisted selection breeding program of germplasm resources, selecting variant germplasm resources meeting a breeding target, and removing germplasm resources which do not meet requirements; then, the branch generating the variation is taken as a scion and grafted on a homologus stock (the homologus stock in the embodiment is 10-year-old 'you-yi' apricot) to perform character fixation until a mutant with expected stable characters is generated;
and (8) expanding the grafted hybrid: starting in the third year after grafting, taking peach flowers with complete growth of grafted hybrid purple-leaf flowering peaches as female parents of pollination units, selecting apricot flower pollen with excellent properties as male parents for pollination and hybridization, wherein the yield of hybrid fruits is improved by more than 3 times compared with that of common hybridization (see table 3), collecting hybrid fruits in the next year, sowing the hybrid fruits and entering a conventional hybridization progeny determination program.
Comparative example 2
Selecting and preparing rootstocks: selecting 'you one' apricots in a plot with the physiological age of 6 years, healthy growth, no plant diseases and insect pests, no gummosis, the plant row spacing of 3m multiplied by 4m, good ventilation and light transmission, higher water and fertilizer conditions and the groundwater level of below 1.5m as rootstocks; the method is implemented in Huang-Huai river basin, 2-4 backbone branches with the thickness of more than 1cm and uniform spatial distribution are selected at a position 1.5m away from the ground of a Youyi apricot stock for short-cutting in 3 months and 5 days, the section is kept smooth and flat, and other branches are completely cut from the base;
selecting scions in the step (2): selecting annual branches as scions, selecting branches of purple-leaf flowering peaches which are annual, have the thickness of 0.6-0.8 cm, full bud eyes, no plant diseases and insect pests and no lateral branches and are positioned at the periphery of a crown, and removing unqualified parts at two ends of the branches to be used as the scions;
step (3), preparation before grafting: weeding and watering the rootstock 5-7 days before grafting, and pretreating the scion; in the embodiment, the selected 'you one' apricot stock land is subjected to weeding and watering once in 3 months and 1 day; before grafting, soaking the scion in clear water until the water level is 2-3 cm higher than the base part of the scion, and soaking for 30-60 min at the ambient temperature of 20-35 ℃;
grafting: grafting the pretreated scion onto a stock to obtain a grafted seedling; the grafting induced variation is carried out by adopting a grafting method, which comprises the following steps: firstly cutting a scion into short branches with the length of 5-7 cm and containing 2-3 buds, cutting the lower ends of the branches into V-shaped branches by using a knife, then cutting the smooth section of a backbone branch to be grafted of a stock into V-shaped notches by using a grafting knife, then embedding the scion into the stock, and binding and compacting the scion by using a plastic film to prevent water from losing; if the thickness of the stock backbone branch is larger than 2cm, a scion can be additionally grafted at the relative position of the first scion, and finally the two scions are fixed by plastic cloth to prevent wind fracture.
Step (5), management after grafting: after grafting, timely preventing and controlling the damage of diseases, pests and weeds, keeping the water content of soil of the grafted seedling at 50-60 wt% after grafting, performing stock shearing and bud picking treatment after the grafting wound is healed, and supplementing exogenous nutrients after the grafted bud grows out; checking once every 7 days after grafting, reserving one stock bud at a position corresponding to the grafted branch, and wiping off other buds; removing sprouts outside the grafted sprouts every 7 days after grafting; when the length of the grafted bud is more than or equal to 10cm, supplementing exogenous nutrients for 1-2 times and keeping soil organic mattersThe content is more than or equal to 1.0 wt% [ organic fertilizer can be applied for 3-5 m per mu of land in dormancy season3Keeping the organic matter content of the soil at 1.0 wt% -5.0 wt%; the exogenous nutrients used were the same as in example 2.
The growth of the grafted scions of example 2 and comparative example 2 is shown in table 2:
TABLE 2 growth of grafted hybrid scion 1 year after grafting by different grafting treatment methods
Note: the significance comparison of the two groups of data adopts Student t test, different letters in the same column represent difference levels, capital letters (A, B) are extremely significant (P is less than or equal to 0.01), and lowercase letters (a and b) are significant (P is less than or equal to 0.05), and the following is the same.
TABLE 3 Effect of different treatments on distant sexual cross-compatibility of grafted hybrids
Note: the significance analysis adopts a Duncan new repolarization difference method, different letters in the same column represent difference levels, capital letters are significant (P is less than or equal to 0.01), and lowercase letters are significant (P is less than or equal to 0.05).
The present inventors summarized and compared the shapes of each grafted hybrid in examples 1 and 2 and comparative examples 1 and 2.
In fig. 1: (a-C) the grafted hybrid 'you-one' apricot + 'purple leaf' flowering peach of example 2, the 'you-one' apricot grafted from the root of comparative example 1, and the grafted hybrid 'purple leaf' flowering peach + 'you-one' apricot of example 1, respectively; (D) the flowers of the grafted hybrid ' you-apricot + ' purple leaf ' flowering peach of example 2 are grayish white; (E) flower of 'you-one' apricot grafted from root in comparative example 1, pink; (F) the flowers of the grafted hybrid 'purple leaf' flowering peach plus 'you one' apricot of example 1 are white; (G-I) is the petal and reproductive organ of the grafted hybrid and the grafted apricot from root corresponding to (A-C), respectively, wherein the yellow arrow indicates the difference in reproductive organ size, petal color and curl; (J-L) is the difference in leaves between the grafted hybrid and the grafted apricot from the root corresponding to (A-C), respectively, and the red arrows show the difference in veins, apexes (back of leaves) and margins; (M) difference in apricot kernel phenotype between the 'you one' apricot grafted from root of comparative example 1 (left) and the grafted hybrid 'you one' apricot + 'purple leaf' flowering peach of example 2 (right); (N) difference in phenotype between the 'you-one' apricot grafted from root of comparative example 1 (left) and the grafted hybrid 'you-one' apricot + 'purple leaf' flowering peach of example 2 (right) almond; (O-P) is the analysis of variance of length, width, thickness and single species of almonds corresponding to (N).
In fig. 2: (a-C) the grafted hybrid 'youyi' apricot + purple leaf 'flowering peach of example 2, the' purple leaf 'flowering peach grafted from root of comparative example 2, and the grafted hybrid' purple leaf 'flowering peach + youyi' apricot of example 1, respectively; (D-F) floral phenotypes corresponding to (A-C), respectively, wherein yellow circles represent floral size and phenotypic differences; (G-I) are the structures of the flowers corresponding to (D-F), respectively, and the yellow lines indicate symmetry; (J-L) is the number of petal-like petals corresponding to (G-I), and yellow arrows indicate the type and number of petal-like flowers. (M-O) are the sepals differences of the corresponding flowers to (J-L), respectively, the yellow circles show sepal size and type differences; (P-R) is the difference of the number of petals corresponding to (M-O), and yellow arrows show the curling condition and the number of the petals; (S-U) are differences of plant leaves corresponding to (M-O), respectively, the yellow circles show the curling state of the leaves, and the red color shows the difference of leaf tips; (V) difference in peach pit phenotype between the 'purple leaf' flowering peach grafted from root of comparative example 2 (left) and the grafted hybrid 'you-one' apricot + 'purple leaf' flowering peach of example 2 (right); (W) is the difference in phenotype of peach kernel in plants corresponding to (V); (X) is a difference analysis of the length, width, thickness and single kernel weight of the peach pit corresponding to (W); (Y) is a difference analysis of peach kernels corresponding to (X). Indicates significant difference (P ≦ 0.01); the scale bar of the background in the figure is 1 cm.
FIGS. 3 and 4 show the growth and development of the grafted bud in example 2 and comparative example 2, respectively, and comparing FIGS. 3 and 4, it can be seen that before grafting, the affinity of grafting can be significantly improved by soaking the scion in a glucanase solution with an enzymatic activity of 100U/L.
Example 3
Selecting and preparing rootstocks: selecting a 'purple leaf' flowering peach seedling which grows in the same year, has a ground diameter (20 cm away from the ground) of more than 0.6cm, is healthy and free of diseases and insect pests and is free of gummosis as a stock in a Huang-Huai river basin for 5 months and 30 days;
selecting scions in the step (2): 6, 7 days in 6 months, selecting an annual branch with 0.5-0.8 cm thickness, plump bud eyes, no plant diseases and insect pests and no side branches at the periphery of a superior-one crown of the kernel apricot variety, and removing unqualified parts of the branch tips to be used as scions;
step (3), preparation before grafting: watering a 'purple leaf' flowering peach stock land once 5-7 days before grafting, and pretreating the scion; in the embodiment, in a shady and cool place with the ambient temperature of 20-35 ℃, the bottoms of branches are aligned, cut, flattened and placed into a scion pretreatment solution for soaking, the soaking solution is 2-3 cm higher than the base part of the scion, the scion pretreatment solution is a melatonin solution with the concentration of 20 mu mol/L, and the solution is soaked for 30-60 min for later use;
grafting: grafting the pretreated scion onto a stock to obtain a grafted hybrid; at the position 5-10cm away from the ground of the purple-leaf flowering peach stock (the ground diameter of the nursery stock at the height is larger than 0.6cm), removing bark to form a rectangular notch on the stock by using parallel double knives with the distance of 0.8-1.0 cm and the distance reaching the xylem, obtaining buds and bud eyes from the 'you first' scion by using the same method, completely attaching the buds and the bud eyes to the rectangular notch of the stock, and finally, tightly binding and consolidating the buds by using a plastic film in a mode of leaking out grafted buds to prevent water from losing; the closer the nursery stock is to the ground, the smaller the physiological age of the nursery stock is; the older the physiological age; for the current-year 'purple-leaf' flowering peach stock, the position with the height of 5-10cm is in the same period with the bud eye obtained from the scion, so that the grafting property is better.
Step (5), management after grafting: on the 7 th day after grafting, the rootstock is cut by using a branch shear 2cm above the grafted bud and the bound plastic strips are removed, so that grafting germination is promoted, wind break is prevented, and a wood stick is required to be arranged in a region with large wind power to protect the bud; after grafting, the water and fertilizer are kept sufficient, and the damage of diseases, pests and weeds is prevented in time; in the last 12 th month, marking the grafted and survived hybrid, planting according to the plant row spacing of 3m multiplied by 5m, and then performing conventional field management;
step (6) inducing variation: in 3 middle of the next month, selecting 1-2 branches with branch angles of 30-60 degrees respectively from new branches sprouting in east, south, west and north directions of a grafting bud and a stock bud respectively, shearing other redundant branches by using branch shears, enabling the grafting bud and the stock bud to form branches with the same biomass, carrying out grafting hybrid induction, and then merging into a conventional field for management;
and (7) fixing characters: starting in the second year after grafting, enhancing water and fertilizer management, removing pulp from the collected grafted hybrid fruits, preserving seed kernels, sowing and entering a germplasm resource determination procedure; meanwhile, the branch generating variation is used as a scion and grafted to a homologus rootstock (in the embodiment, the homologus rootstock is the current-year ' purple-leaf ' flowering peach ') to perform character fixation until a mutant with expected stable characters is generated;
and (8) expanding the grafted hybrid: starting in the third year after grafting, taking the apricot flowers with complete development of grafted hybrids 'you one' as a pollination unit female parent, selecting 'purple leaf' flowering peach pollen with excellent characters as a male parent for pollination and hybridization, wherein the yield of hybrid fruits is improved by more than 2.3 times compared with common hybridization, and collecting hybrid fruits in the next year for sowing and entering a conventional hybridization progeny determination program.
Comparative example 3
Selecting and preparing rootstocks: selecting a 'purple leaf' flowering peach seedling which grows in the same year, has a ground diameter (20 cm away from the ground) of more than 0.6cm, is healthy and free of diseases and insect pests and is free of gummosis as a stock in a Huang-Huai river basin for 5 months and 30 days;
selecting scions in the step (2): 6, 7 days in 6 months, selecting an annual branch with 0.5-0.8 cm thickness, plump bud eyes, no plant diseases and insect pests and no side branches at the periphery of a superior-one crown of the kernel apricot variety, and removing unqualified parts of the branch tips to be used as scions;
step (3), preparation before grafting: watering the stock land once 5-7 days before grafting, and pretreating the scion; in the embodiment, the bottom of a branch is aligned, cut, leveled and placed into clear water to be soaked in a shade place with the environment temperature of 20-35 ℃, the soaking solution is 3cm higher than the base part of the scion, and the soaking time is 30-60 min for later use;
grafting: grafting the pretreated scion onto a stock to obtain a grafted hybrid; the method comprises the following steps of (1) removing barks of purple-leaf flowering peach stocks 5-10cm away from the ground by using parallel double knives with the distance of 0.8-1.0 cm on the stocks to form rectangular gaps, obtaining buds and bud eyes from 'you first' scions by using the same method, completely attaching the buds and the bud eyes to the rectangular gaps of the stocks, and finally, tightly binding and compacting by using plastic films in a mode of leaking grafted buds to prevent water from losing;
step (5), management after grafting: on the 7 th day after grafting, the rootstock is cut by using a branch shear 2cm above the grafted bud and the bound plastic strips are removed, so that grafting germination is promoted, wind break is prevented, and a wood stick is required to be arranged in a region with large wind power to protect the bud; after grafting, the water and fertilizer are kept sufficient, and the damage of diseases, pests and weeds is prevented in time; in the last 12 th month, marking the grafted and survived hybrid, planting according to the plant row spacing of 3m multiplied by 5m, and then performing conventional field management;
and (6): manually erasing the germinated branches except for the grafted branches every 10-15 days in the middle ten days of the 3 rd month of the next year, and then merging the germinated branches into the conventional field management.
FIGS. 5 and 6 are the growth and development of the grafted bud in the second 4 months of the second year after grafting in example 3 and comparative example 3, respectively, in FIG. 5, the rootstock and the scion grow basically in the same way, and in FIG. 6, the rootstock and the scion grow in different ways to form a small foot; comparing fig. 5 and fig. 6, it can be seen that, before grafting, the grafting scion is soaked in the melatonin solution with the concentration of 20 μmol/L, which is beneficial to significantly improving the grafting affinity.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications are possible which remain within the scope of the appended claims.
Claims (10)
1. The method for mutagenesis of peach-apricot intergeneric grafting hybrid is characterized by comprising the following steps:
selecting and preparing rootstocks: selecting apricot plants or peach plants as rootstocks;
selecting scions in the step (2): selecting annual or current-year branches as scions; the scion is a peach plant or apricot plant and is of a different species from the stock;
step (3), preparation before grafting: before grafting, weeding and watering the rootstock, and pretreating the scion;
grafting: grafting the pretreated scion onto a stock to obtain a grafted seedling;
step (5), management after grafting: keeping the water content of the soil of the grafted seedling at 50-60 wt% after grafting, performing stock shearing and bud picking treatment after the grafting wound is healed, and supplementing exogenous nutrients after the grafted bud grows out;
step (6) inducing variation: controlling the biomass ratio of the rootstock and the scion by a manual pruning mode to induce the character variation of the grafted seedling to obtain a grafted hybrid;
and (7) fixing characters: fixing the character of the branch with excellent character variation by sowing, grafting or tissue culture;
and (8) expanding the grafted hybrid.
2. The method for mutagenesis of peach-apricot intergeneric grafting hybrids according to claim 1, wherein in step (1), the apricot plant is common apricot (a. vulgaris), siberian apricot (a. sibirica), Liaoning apricot (a. mandshurica), Tibetan apricot (a. holosecia), purple apricot (a. dasycarpa), Shidanapricot (a. zhidannensis), plum (a.plum), plum apricot (a.limeixing), political apricot (a.zhenghensis) or Chinese almond apricot (a.cathayana d.l.fu, b.r.li j.h.li, sp.nov.); the Prunus plant is Prunus (Amygdalus L.) or Prunus Amygdalus (subg. Persica L.); the subgenus persicae is fructus Caricae (A.davidiana), Sinkiang fructus Persicae (A.ferganensis), Gansu fructus Persicae (A.kansuensis), semen Juglandis Mandshuricae (A.mira) or fructus Persicae (A.persica); amygdalus belongs to amygdalus, and combinations thereof, and amygdalus.
3. The method for mutagenesis of peach-apricot intergeneric grafting hybrid according to claim 1, characterized in that in step (1), peach trees or apricot trees which grow healthy and have no plant diseases and insect pests and no gummosis are selected as rootstocks, and the row spacing of the peach trees or apricot trees is (2-3) mx (4-8) m;
if grafting in spring: selecting a trunk or a backbone branch with the rootstock thickness of more than or equal to 1cm, cutting the trunk or the backbone branch at a position 0.4-1.5 m away from the ground, and cutting the section into a smooth plane for later use;
if grafting in summer: in spring, the trunk or the bone trunk branches with the thickness of more than or equal to 1cm are cut, the current-year new shoots with the thickness of more than 0.6cm are selected as the grafting-planning objects of the scions, and other branches which do not meet the standard are removed.
4. The method for mutagenesis of peach-apricot intergeneric grafting hybrid according to claim 1, characterized in that in step (2), an annual or current annual branch without side branches located at the periphery of crown and having thickness of 0.5-1.0 cm is selected, and the part of the branch with unqualified tip is removed to be used as scion;
in the step (3), watering the rootstock thoroughly 5-7 days before grafting;
in the step (4), grafting is carried out in a mode of grafting by adopting a branch grafting mode or a wood bud grafting mode in spring grafting, and grafting is carried out in a mode of square bud grafting or a wood bud grafting mode in summer grafting; the grafting time in spring is before the spring equinox after rainwater, and the grafting time in summer is before the summer late summer of the miscanthus.
5. The method for mutagenesis of peach-apricot intergeneric grafting hybrids as claimed in claim 5, characterized in that the spring grafting is performed by means of grafting: cutting the scion into short branches containing 2-3 buds and 5-7 cm in length, cutting the lower end of the scion into a V shape by using a grafting knife, simultaneously cutting a smooth plane of a grafting part of the stock into a corresponding V-shaped notch, or vertically cutting a wound with the depth of 2-3 cm downwards on one side of the grafting part of the stock, and then embedding the scion into the stock; binding and compacting the stock and the scion by using a plastic film;
grafting in summer by adopting a square bud grafting mode: selecting current-year branches with the thickness of 0.6-1.0 cm from the rootstocks as grafting objects of the scions, and removing barks on the rootstocks and reaching xylem deeply by using parallel double knives with the distance of 0.8-1.0 cm to form rectangular gaps; and (3) obtaining buds and bud eyes from the scion by the same method, completely attaching the buds and the bud eyes to the rectangular notch on the stock, tightly binding and compacting by using a plastic film, and enabling grafted buds to leak out and expose out of the plastic film.
6. The method for mutagenesis of peach-apricot intergeneric grafting hybrid according to claim 1, wherein in the step (3), the pretreatment method of the scion is: immersing 2-3 cm of a scion base part into the scion pretreatment liquid, and immersing for 30-60 min at the temperature of 20-35 ℃; the scion pretreatment solution is a melatonin solution, or a glucanase solution, or a mixed solution of the melatonin solution and the glucanase solution; the concentration of the melatonin solution is 10-20 mu mol/L, and the active concentration of the dextranase solution is 50-100U/L.
7. The method for mutagenesis of peach-apricot intergeneric grafting hybrid according to claim 1, characterized in that in step (5), after 3 days of grafting, anvil folding is performed at a distance of 3-5 cm from the grafted bud; cutting stocks 1-2 cm above the grafting openings 7-10 days after summer grafting or 25-30 days after spring grafting, and removing the binding objects for grafting; erasing other sprouts on the rootstock except for the grafting sprouts; when the length of the grafted bud is more than or equal to 10cm, supplementing exogenous nutrients for 1-2 times, and keeping the organic matter content of the soil to be more than or equal to 1.0 wt%; in the exogenous nutrients, the mass ratio of nitrogen to phosphorus to potassium is 8: 3: 6.
8. the method for mutagenesis of peach-apricot intergeneric grafting hybrid according to claim 1, characterized in that in step (6), the biomass ratio of the new branch of stock germination and the scion grafting bud is controlled in 1: (0.5-4) to induce the character variation of the grafted seedling.
9. The method for mutagenesis of peach-apricot intergeneric grafting hybrid according to claim 1, wherein in the step (7), the method for fixing the character and enhancing the variation comprises: collecting the fruit of the grafted hybrid, removing the pulp, and reserving the seed core for sowing;
or, taking the branch generating variation as a scion, and grafting the scion to the same stock for character fixation;
or, the branch generating the variation is taken as the scion and grafted to the xenogeneic rootstock, and the germination of the xenogeneic rootstock is completely limited;
or, carrying out asexual propagation culture by using the variant-producing branches as explants to obtain complete plants.
10. The method for mutagenesis of peach-apricot intergeneric grafting hybrid according to claim 1, wherein in step (8), pollen of peach or apricot is collected and artificial pollination is performed with the grafting hybrid after the third year and stable character of the grafting hybrid.
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