阅读说明：本技术 一种可以缩短育种周期的番木瓜杂交后代成年结果树潜伏芽萌发方法及其应用 (Papaya filial generation adult fruiting tree latent bud germination method capable of shortening breeding period and application thereof ) 是由 魏岳荣 杨敏 周陈平 邝瑞彬 杨护 黄炳雄 于 2021-07-30 设计创作，主要内容包括：本发明属于植物育种技术领域,具体涉及一种可以缩短育种周期的番木瓜杂交后代成年结果树潜伏芽萌发方法及其应用,本发明用GA-(3)+6-BA混合液喷施番木瓜杂交后代成年结果树结果部位以下的主干潜伏芽位点至滴液,7-15天后重复喷施第二次,第二次喷施7-15天后重复喷施第三次,第三次喷施后生长20天即可获得1cm以上的侧芽,获取的侧芽可以用于作为组织培养快繁的外植体进行种苗繁育。本发明方法可对正处于结果期的杂交后代优株提前进行种苗繁育,既避免了传统育种方法中通过截干来获得侧芽而影响综合性状评价,又能避免杂交F-(1)代过冬易感染环斑花叶病毒病从而无法获得优株后代的风险,从而大大缩短了育种周期。(The invention belongs to the technical field of plant breeding, and particularly relates to a papaya filial generation adult fruiter latent bud germination method capable of shortening a breeding period and application thereof 3 And spraying the main latent bud site below the fruiting part of the papaya hybrid progeny fruit tree to dropping liquid in the mixed liquid of +6-BA, spraying for the second time after 7-15 days, spraying for the third time after 7-15 days, and growing for 20 days after the third spraying to obtain the lateral bud of more than 1cm, wherein the obtained lateral bud can be used as an explant for tissue culture and rapid propagation to carry out seedling breeding. The method can be used for advancing the excellent hybrid plants in the fruiting periodSeedling breeding avoids influencing comprehensive character evaluation by obtaining lateral buds through stem cutting in the traditional breeding method and avoids cross breeding F 1 The overwintering is susceptible to the ringspot mosaic virus, so that the risk that excellent plant progeny cannot be obtained is avoided, and the breeding period is greatly shortened.)

1. A method for promoting germination of latent buds of adult fruiter trees of papaya filial generation is characterized in that GA is used₃And spraying hormone mixed solution of 6-BA on the trunk latent bud site below the fruiting part of the papaya filial generation adult fruiting tree to dropping liquid, repeatedly spraying for the second time after 7-15 days, repeatedly spraying for the third time after 7-15 days, and growing for 20 days after the third spraying to obtain lateral buds with the length of more than 1 cm.

2. The method for promoting germination of latent buds of adult bearing trees of papaya filial generation according to claim 1, wherein the adult bearing trees of papaya filial generation are papaya cross F₁Generation of superior plants and fruit trees.

3. The method for promoting germination of latent buds of adult fruiting trees of papaya hybrid offspring according to claim 2, wherein the papaya hybrid F is₁The substitute excellent strain comprises 'GZ 16-8' papaya, 'GZ 16-4' papaya and 'GZ 20-1' papaya.

4. The method of claim 1, wherein GA is added to the hormone mixture to promote germination of the latent bud of the mature fruiter tree of the papaya filial generation₃The concentration of (b) is 200-600mg/L, and the concentration of 6-BA is 100-300 mg/L.

5. The method as claimed in claim 4, wherein GA is added into the hormone mixture solution₃The concentration of (A) is 400mg/L, and the concentration of 6-BA is 200 mg/L.

6. The method for promoting the germination of the latent buds of the adult fruiter trees of the papaya filial generation according to claim 1, wherein the spraying of the hormone mixed solution is performed in sunny weather.

7. The method for promoting the germination of the latent buds of the adult fructification trees of the papaya hybrid progeny as claimed in any one of claims 1 to 6 is applied to rapid propagation and seedling cultivation of papaya tissue culture.

Technical Field

The invention belongs to the technical field of plant breeding, and particularly relates to a papaya filial generation adult fruiter latent bud germination method capable of shortening a breeding period and application thereof.

Background

Papaya (Caricapapaya L.) is a large perennial herbaceous fruit tree with evergreen and soft wood in tropical and subtropical zones, is well known as the good fruit in Lingnan, has the nutritional value of the first ten nutritional fruits in the world, and is an important economic crop for adjusting the agricultural variety structure in China.

The traditional breeding mode of the papaya is cross breeding, for example, Chinese patent CN201710946695 discloses a papaya cross breeding method, which comprises the steps of parent selection, pollen preparation, emasculation, pollination, field management, hybrid seed harvesting, seed sowing, hybrid progeny optimization, evaluation and the like. Due to the fact that the number of the papaya seeds is large, the character separation of the later generations is obvious, the multi-generation screening and evaluation workload of the filial generations is large, the difficulty is large, and the cost is high, the breeding period is long, and the breeding efficiency is low. Therefore, how to improve the optimization and evaluation work efficiency of the filial generation and shorten the breeding period becomes the important research point of the papaya cross breeding.

For papaya F₁The serious character separation of the generation seed seedlings applies the tissue culture rapid propagation technology to the cultivation of the hybrid F₁The asexual propagation seedling of the generation excellent plant (single plant with excellent comprehensive characters) can not only keep the genetic stability of the excellent plant and avoid the phenomenon of the separation of the characters of the generation after the seedling of the seed, but also can cultivate a large amount of seedlings in a short time for the comprehensive character evaluation and variety test of filial generation, thereby greatly shortening the breeding period and improving the breeding efficiency. Regarding the tissue culture and rapid propagation technology of papaya, the Chinese invention patent CN1202706C discloses a method for producing papaya with stable plant propertyA method for tissue culture of pawpaw seedlings; CN108112479B discloses a method for raising the roots of papaya tissue culture seedlings by stem section suspension leaf planting of differentiated buds. The two methods are implemented by cutting off the trunk of the adult fruit tree, naturally germinating the lateral bud to obtain the explant for tissue culture, and performing disinfection, bud proliferation and rooting to obtain the tissue culture seedling. If the above two methods are directly applied to papaya hybridization F₁The seedling breeding of the generation excellent plant needs to be firstly carried out with the stem cutting treatment. In the conventional papaya cross breeding process, the character evaluation of a complete breeding cycle of the excellent plants of the filial generation is usually required, and the lateral buds can be obtained for seedling propagation after the trunk cutting in the second year after the evaluation. As papaya plants cultivated in a field are very easy to infect the ringspot mosaic virus and even die after the low temperature of the field in winter, F cannot be obtained₁Great risk of offspring of generation-excellent plants. Therefore, if the hybrid F can be broken by external force in the stage of full-length of the result without cutting the stem and without affecting the evaluation of the integrity of the excellent strain₁Promoting the germination of latent buds of the trunk part by the top advantages of the generation excellent plant adult fruiting tree, and obtaining F by utilizing the tissue culture rapid propagation technology₁The asexual propagation seedling of the generation excellent plant is applied to variety tests, and the breeding period can be further shortened. Therefore, a method for promoting the germination of the latent buds of the adult fruiting trees of the papaya hybrid progeny is needed to accelerate the cross breeding process of the papaya.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention provides a method for promoting the germination of latent buds of adult fruiting trees of papaya hybrid progeny, which is applied to tissue culture rapid propagation (short for tissue culture rapid propagation) and seedling culture of excellent plants of papaya hybrid progeny and can greatly shorten the breeding period, thereby effectively solving the problems that the existing papaya hybrid progeny is susceptible to the ringspot mosaic virus disease after overwintering, the excellent plant progeny cannot be obtained, the cross breeding period is long, the evaluation workload is large and the efficiency is low.

In order to achieve the purpose, the invention adopts the technical scheme that:

papaya filial generation adult knot promoting papaya filial generationThe method for germinating the latent buds of the fruit trees comprises the following specific steps: by GA₃The hormone mixed solution of (gibberellin) and 6-BA (benzylamino adenine) is sprayed to main latent bud sites below fruiting parts of adult fruiting trees of papaya filial generations to dropping liquid, the spraying is repeated for the second time after 7-15 days, the spraying is repeated for the third time after 7-15 days of the second spraying, and lateral buds with the length of more than 1cm can be obtained after the lateral buds grow for 20 days after the third spraying, so that the lateral buds can be used as explants for tissue culture and rapid propagation.

Auxin produced by plant terminal buds is transported downwards due to apical dominance to inhibit the growth of lateral buds, so that the lateral buds of the papaya plants in the fruiting period can not germinate, cytokinin can remove apical dominance caused by the auxin to promote the growth and development of the lateral buds, and gibberellin GA₃Cell elongation and cell division can be promoted, so that the present invention uses 6-BA to relieve apical dominance and uses GA₃To promote the elongation growth of the germinating lateral buds.

Preferably, the adult fruiting tree of the papaya filial generation is papaya cross F₁Generation of superior plants and fruit trees.

Further, the papaya hybrid F₁The substitute excellent strain comprises 'GZ 16-8' papaya, 'GZ 16-4' papaya and 'GZ 20-1' papaya.

Preferably, in the hormone mixture, GA₃The concentration of (b) is 200-600mg/L, and the concentration of 6-BA is 100-300 mg/L.

Further, in the hormone mixture, GA₃The concentration of (A) is 400mg/L, and the concentration of 6-BA is 200 mg/L.

Preferably, the spraying of the hormone mixed solution is carried out in sunny weather.

The invention also provides application of the method for promoting the germination of the latent buds of the adult fructification trees of the papaya hybrid progeny in tissue culture and rapid propagation and seedling culture of papaya.

Compared with the prior art, the invention has the beneficial effects that:

the invention provides a method for promoting germination of latent buds of adult fruiting trees of papaya filial generations, which uses GA₃+6-BA mixed liquor is sprayed on the fruit bearing part of the papaya hybridization progeny adult fruiting treeThe germination of the main latent bud of the fruiting tree can be promoted to start to grow from the position of the dry latent bud to the dropping liquid, the spraying is repeated for the second time after 7-15 days, the spraying is repeated for the third time after 7-15 days, the side bud with the length of more than 1cm can be obtained after the growth of 20 days after the spraying is performed for the third time, and the obtained side bud can be used as an explant for tissue culture and rapid propagation to breed seedlings. Because the method can cultivate asexual propagation seedlings for variety test by the tissue culture rapid propagation technology in advance for the excellent hybrid progeny plants in the fruiting period, the method avoids the influence on comprehensive character evaluation caused by the lateral buds obtained by stem cutting in the traditional method and also avoids the cross F₁The generation overwintering is susceptible to the ringspot mosaic virus disease, so that the risk of obtaining excellent plant progeny cannot be obtained, meanwhile, a large number of excellent plant progeny with stable properties and good consistency can be obtained by the method for variety test, the breeding period can be effectively shortened, and the problems of long period, large evaluation workload and low efficiency of the conventional papaya cross breeding are solved.

Drawings

FIG. 1 shows the whole process of latent bud germination of hybrid F1 generation superior plant 'GZ 16-8' in full bearing stage (A is the stem without lateral bud in the first spraying treatment, B is the stem 10 days after the first spraying treatment and with a small amount of lateral bud germination, C is the stem in the third spraying treatment and with a small amount of lateral bud germination, D is the stem in the harvesting stage 20 days after the third spraying treatment and with the germinated lateral bud for tissue culture and rapid propagation);

FIG. 2 is the stem of hybrid F1 generation superior plant 'GZ 16-4' plant in harvest time after the third spraying treatment and after growing for 20 days (the germinated lateral bud can be used for tissue culture and rapid propagation).

Detailed Description

The following further describes the embodiments of the present invention. It should be noted that the description of the embodiments is provided to help understanding of the present invention, but the present invention is not limited thereto. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.

The experimental procedures in the following examples were carried out by conventional methods unless otherwise specified, and the test materials used in the following examples were commercially available by conventional methods unless otherwise specified.

Example 1 method for promoting germination of latent buds of adult fruiting trees of papaya filial generations

The papaya hybrid progeny adult fruiting tree of this example is papaya hybrid F₁The generation excellent plant GZ16-8 fruit bearing tree has the flow of promoting the germination of latent buds as follows: the latent bud of the fruit bearing trunk → the spraying of the hormone mixed liquid → the repeated spraying of the 2 nd time after 10 days → the repeated spraying of the 3 rd time after 10 days → the growth of the lateral bud for 20 days → the ideal lateral bud. The specific method for promoting the latent bud germination comprises the following steps:

(1) selecting 400mg/L GA for sunny weather₃The mixture of +200 mg/L6-BA was sprayed to the stem latent bud site below the fruiting site (the stem when sprayed for the first time is shown in FIG. 1A) to the dropping liquid, the latent bud was seen to germinate and grow 7 days later, and the stem 10 days later is shown in FIG. 1B.

(2) The spraying is repeated for the second time after 10 days of the first spraying.

(3) The spraying was repeated a third time 10 days after the second spraying, and the shoots at the time of the third spraying treatment were as shown in FIG. 1C.

(4) After the lateral bud is sprayed for the third time, the lateral bud with the length of more than 1cm can be obtained after the lateral bud grows for 20 days (as shown in figure 1D), and the lateral bud can be used as an explant for tissue culture and rapid propagation to carry out seedling breeding.

Example 2 method for promoting germination of latent buds of adult fruiting trees of papaya filial generations

The papaya filial generation adult fruity tree of this example is 'GZ 16-4' papaya cross F₁The process of promoting the germination of latent buds of a generation-superior plant fruiting tree generally comprises the following steps: the latent bud of the fruit bearing tree stem → the spraying of the hormone mixed liquid → the repeated spraying of the 2 nd time after 7 days → the repeated spraying of the 3 rd time after 15 days → the growth of the lateral bud for 20 days → the ideal lateral bud. The specific method for promoting the latent bud germination comprises the following steps:

(1) selecting 400mg/L GA for sunny weather₃+200mg/L of 6-BA mixed solution is sprayed to the position of the latent bud of the main stem below the fruiting part until dropping liquid, and the latent bud can germinate and grow after 7 days.

(2) The spraying is repeated for the second time 7 days after the first spraying.

(3) The spraying is repeated for the third time after 15 days of the second spraying.

(4) After the lateral bud is sprayed for the third time, the lateral bud with the length of more than 1cm can be obtained after the lateral bud grows for 20 days (as shown in figure 2), and the lateral bud can be used as an explant for tissue culture and rapid propagation to carry out seedling breeding.

Example 3 method for promoting germination of latent buds of adult fruiting trees of papaya filial generations

The papaya filial generation adult fruity tree of this example is 'GZ 20-1' papaya cross F₁The process of promoting the germination of latent buds of a generation-superior plant fruiting tree generally comprises the following steps: the latent bud of the fruit bearing tree stem → the spraying of the hormone mixed liquid → the repeated spraying of the 2 nd time after 15 days → the repeated spraying of the 3 rd time after 7 days → the growth of the lateral bud for 20 days → the ideal lateral bud. The specific method for promoting the latent bud germination comprises the following steps:

(1) selecting 400mg/L GA for sunny weather₃+200mg/L of 6-BA mixed solution is sprayed to the position of the latent bud of the main stem below the fruiting part until dropping liquid, and the latent bud can germinate and grow after 7 days.

(2) The spraying is repeated for the second time after 15 days of the first spraying.

(3) The spraying is repeated for the third time after 7 days of the second spraying.

(4) After the lateral bud is sprayed for the third time, the lateral bud grows for 20 days to obtain the lateral bud with the length of more than 1cm, and the lateral bud can be used as an explant for tissue culture and rapid propagation to carry out seedling breeding.

Example 4 method for promoting germination of latent buds of adult fruiting trees of papaya hybrid progeny

The specific method is the same as example 1, except that the mixed solution sprayed in the step (1) is 200mg/L GA₃+100mg/L of 6-BA, and the subsequent repeated spraying uses the mixed liquid with the concentration.

Example 5 method for promoting germination of latent buds of adult fruiting trees of papaya hybrid progeny

The specific method is the same as example 1, except that the mixed solution sprayed in step (1) is 600mg/L GA₃+300mg/L of 6-BA, and subsequent repetitionThe mixed solution with the concentration is used for spraying.

Comparative example 1 method for promoting germination of latent buds of adult fruiting trees of papaya filial generations

The specific method is the same as the example 1, except that the third spraying process of the step (3) is omitted.

Comparative example 2 method for promoting germination of latent buds of adult fruiting trees of papaya filial generations

The specific method is the same as the example 1, except that the second spraying process and the third spraying process in the steps (2) and (3) are simultaneously lacked.

Comparative example 3 method for promoting germination of latent buds of adult fruiting trees of papaya filial generations

The specific method is the same as the example 1, except that the step (2) is repeated spraying for a second time after 20 days of first spraying.

Comparative example 4 method for promoting germination of latent buds of adult fruiting trees of papaya filial generations

The specific method is the same as the example 1, except that the step (3) is repeated spraying for the third time after 20 days of spraying for the second time.

Comparative example 5 method for promoting germination of latent buds of adult fruiting trees of papaya filial generations

The specific method is the same as the example 1, except that the step (2) is repeated spraying for the second time after 5 days of first spraying.

Comparative example 6 method for promoting germination of latent buds of adult fruiting trees of papaya filial generations

The specific method is the same as the example 1, except that the step (3) is repeated spraying for the third time after 5 days of spraying for the second time.

The method for promoting the germination of the latent buds of the adult fruit trees of the papaya filial generations in the examples 1-5 and the comparative examples 1-6 is subjected to implementation effect statistics, and the statistical items comprise the initial germination time, the average germination number of a single plant and the number of healthy lateral buds of more than 1cm which can be obtained after the lateral buds grow for 20 days after the third spraying treatment. The statistical results are shown in table 1.

As can be seen from the results in Table 1, using the method for promoting germination of latent buds according to the invention, hybrid F of different hybrid combinations₁The generation excellent plants can obtain 12-14 germinated latent buds, the number of lateral buds with the length of more than 1cm which can be obtained after the lateral buds grow for 20 days after the third spraying treatment is 6-8, and the mixed solution after the spraying treatment is 200-600mg/L GA-₃+100-300 mg/L6-BA obtained more than 1cm lateral bud by the treatment combination of example 1 (400mg/L GA)₃+200mg/L of 6-BA mixed liquor) has the highest sprouting quantity and lateral bud quantity above 1cm, but is lack of secondary spraying or/and third spraying; the second spraying time is too early or too late; if the third spraying time is too early or too late, lateral buds above 1cm cannot be obtained or only a small amount of lateral buds above 1cm can be obtained.

Therefore, the method can be used for obtaining the explant used for tissue culture and rapid propagation to carry out seedling propagation. At the same time, the method of the present invention can align the hybrid F in the fruiting period₁The generation excellent plants are used for cultivating asexual propagation seedlings by the tissue culture rapid propagation technology in advance, so the breeding period can be effectively shortened, the comprehensive character evaluation cannot be influenced by stem cutting, and the cross F can be avoided₁The generation needs to pass winter and is susceptible to the ringspot mosaic virus, so that the risk that excellent plant progeny cannot be obtained is caused, and the breeding efficiency is greatly improved.

TABLE 1 results of promoting germination of latent buds of adult bearing trees of different papaya filings generations

The embodiments of the present invention have been described in detail, but the present invention is not limited to the described embodiments. It will be apparent to those skilled in the art that various changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, and still fall within the scope of the invention.