CN112544366A - Cultivation method of passion fruit seedlings - Google Patents

Abstract

The invention belongs to the field of high-quality seedling cultivation and application, and relates to a cultivation method of passion fruit seedlings. The method creatively selects the filial generation F1 obtained by hybridizing passion flower with fine teeth and passion flower with green fruit with stronger resistance as the stock, selects the tender branch of passion fruit with leaf buds as the scion, and the seedlings cultivated by the grafting method have strong resistance to the stem rot and luxuriant branches of the seedlings, thereby reducing the updating planting frequency of the seedlings in the planting process of the passion fruit variety, reducing the cultivation and management cost, prolonging the service life of the passion fruit plant and increasing the harvesting age of the fruit.

Description

Cultivation method of passion fruit seedlings

Technical Field

The invention belongs to the field of high-quality seedling cultivation and application, and particularly relates to a cultivation method of passion fruit seedlings.

Background

Currently, the edible variety of passion flower is mainly 6 types: passion fruit, passion fruit (passsiflora edulis Sims), passion fruit (p.edulis sim.f. flavicarpa Deg.), passion fruit (p.quadratus), passion fruit (p.malalissima, Kunth) l.h.bailey, passion fruit with camphor (p.laurifolia L.), passion fruit (p.ligularis Juss); the horticultural varieties of purple and yellow fruit species and the series of varieties that are hybridized with each other are customarily called passion fruit. The passion fruit is a tropical perennial evergreen herbaceous to woody climbing vine plant of passion of Passifloraceae, the origin is located in Australia and Brazil, the passion fruit is widely planted in tropical and subtropical regions, and the passion fruit is introduced and cultivated as a fruit crop in China, and the year is nearly three or forty years.

The passion fruit stem is cylindrical and has fine stripes and no hair; the leaf is paper, the length is 6-13 cm, the width is 8-13 cm, the palm is deep-cracked or not split, the base part of the near-cracked piece is provided with 1-2 cupular glands, no hair is left, and the base part has tendrils; flower fragrance, diameter 6-10 cm; 5 sepals are obtained; 5 petals which are as long as sepals; the outer auxiliary corolla splinters are 4-5 rounds, and the outer 2 rounds are filiform, and are approximately equal to the petals in length; berry is egg-shaped, has a diameter of 5-8 cm, and has no hair. The flowering period is 3-9 months, and the fruit period is 6-12 months. At present, the main cultivated varieties in the market are gypsophila paniculata, Zixiang, Tainong, gold fruit and the like.

The passion fruit juice has rich fragrance and proper sour and sweet taste, and can be used as fresh fruit or made into beverage. The passion fruit is rich in nutrient components, amino acids, vitamins, mineral elements, alkaloid, flavonoid and the like; it has multiple health promotion functions, such as promoting fluid production to quench thirst, relieving fatigue, caring skin, inhibiting bacteria, resisting cancer, and reducing cholesterol. In addition, the passion fruit has large and gorgeous flowers, long flowering phase and easy cultivation, is an ideal plant for arranging flower walls and hedges in a garden, and is a fruit crop with high economic value integrating edible, medicinal and ornamental functions.

Since the introduction, the passion fruit has the advantages of high economic value, easiness in planting management and the like, so that the passion fruit is widely concerned, the planting area is increased year by year, the passion fruit is widely planted in tropical and subtropical regions of China, and the planting areas are mainly concentrated in Taiwan, Guangdong, Fujian, Guangxi, Yunnan, Zhejiang, Sichuan and the like. The types of the passion fruit planted in large area in China are mainly purple fruit series, but the purple fruit series have genetic defects susceptible to stem basal rot. The incidence rate of the stem base rot of the passion fruit rooting seedlings in the second year in southern and southwest planting areas and eastern planting areas of the passion fruit can reach 100%. The field onset symptoms of the disease are as follows: the cortex of the basal part of the stem of the infected plant becomes dark, decays and breaks away from xylem, and the leaves wither and fall off. The plant can still grow when only part of cortex of the basal part is infected, and a rotten circle is formed when all cortex is infected, so that the plant gradually dies. The rotten parts are mainly concentrated at the base of the stem 5-10 cm from the ground, and can grow upwards to the cortex of the stem 10-20 cm from the ground and downwards to the root system. When the weather is wet, white mycelium and orange capsula can be seen on the surface of the infected plant tissue. The disease mainly occurs in high-temperature and rainy seasons, and is commonly found in the stem base of plants with the seedling age of more than 1 year.

The stem-root rot is one of the main diseases frequently occurring in the process of planting passion fruits in various countries, and the main pathogenic bacteria of the stem-root rot are different due to different regions, for example, the main pathogenic bacteria in the Fujian area are Fusarium oxysporum (Fusarium oxysporum), the Hainan Qinhuan is Phytophthora nicotianae (Phytophthora nicotianae), the Taiwan is Fusarium solani (F.solani), the Wuganda is Fusarium solani (F.solani), the New Zealand Ox area is Fusarium sambucinum (F.sambucinum), so that the main pathogenic bacteria of the stem-root rot of the passion flower in the Western Banana area can not be inferred through the main pathogenic bacteria reported in other areas. Separating and purifying diseased parts of diseased plants by a traditional separation method to obtain strains, finding out and determining main pathogenic strains by living inoculation and the Koehsch's rule, and determining that the main pathogenic bacteria of the passion fruit stem base rot in the Xishuangbanna area is Fusarium solani (F.solani), the front of the colony of the pathogenic bacteria on a PDA culture medium is white villous, the back of the colony is cream yellow, and the colony has small conidia of egg shape, ellipsoid shape or kidney shape, 1 or no transverse septum and the length of 7.98-21.64 um; the large conidiophore is straight or slightly curved, is shaped like a sickle, has 3 or 5 transverse septa and is 22.52-46.67um long. The spores have strong viability and are very difficult to inactivate, can survive and overwinter in diseased plants, diseased residues or soil in the field, and can propagate and infect the plants by wind, rain and irrigation water when the environmental conditions are proper.

Passiflora incarnata (Passiflora serrulata Jacq.) has dark green stem, cylindrical shape, hollow shape, dense hair, leaf paper shape, heart shape, two or three shallow cracks or no split, length of 13-17 cm, width of 12-14 cm, 1 pair of small cupular gland near the base of leaf stalk, no hair, and twining beard and flower. The diameter of the flower is 5-8 cm; 3 bracts, light green; 5 sepals with the length of 4-5 cm; 5 petals, light green bottom color and dark purple spots in the middle; the external corolla is purple red, white alternate stripes and threadlike; berries are approximately spherical, 2-4cm in diameter, and orange in color at maturity. According to the field growth condition and the laboratory living body inoculation verification, the passion flower with serrate teeth has stronger capability of resisting basal rot than the passion flower with purple fruits.

Passion fruit (passsiflora maliformis (Kunth) l.h.bailey) is cylindrical in stem, hollow, hairless, leathery, dark green, long-distance circular, 12-15 cm long, 5-8 cm wide, with micro-serrations at the leaf margin, 1 pair of cupular glands in the leaf stalk from the base 1/2, hairless, with tendrils. The diameter of the flower is 8-10 cm; 3 bracts, 6-7 cm long, 3-4 cm wide and light yellow green; 5 sepals are obtained; 5 petals; 2 external corolla wheels, filamentous shape, red, purple and light purple alternate stripes; berry is egg-shaped, unhaired, light green in peel, with small white spots, thicker and harder, and white green in juice.

The filial generation of the green dentate (p.serilato p.maliformis, and p.maliformis p.serilato) has a cylindrical stem, is hollow, hairless, has a cutin, is dark green, is round in long distance, has two or three shallow fissures or is not fissured, has a length of 15-20 cm and a width of 12-16 cm, and has 1 pair of cupular glands at the position of a leaf stalk from a base 1/2; the flower diameter is about 6 cm; 3 bracts, light green, 6-8 cm long and-4 cm wide; the sepals are 5 pieces, the length is 4-5 cm, and the width is 1-2 cm; the flower petal is 5cm long and 0.5-1 cm wide, and the flower petal is 2 rounds of external corolla, is threadlike and has red, purple and light purple alternate stripes; berry egg-shaped; the phenotype characters of filial generations of the serrate green fruit are very similar, the phenotypic characters of the filial generations of the serrate green fruit are all synthesized with the morphological characteristics of a male parent and a female parent, the material and the color of leaves are similar to those of passion flower of the green fruit, the shape and the size are similar to those of the passion flower of the serrate green fruit, and the filial generations have two to three shallow cracks or are not split; the flower color is similar to that of passion fruit, and the stigmatic spot characteristic is similar to that of passion fruit with serrate; the fruit size is between the parents, the appearance is similar to that of passion fruit, and the color at maturity is similar to that of passion fruit. Moreover, the filial generation of the green dentate has obvious heterosis and very strong growth potential, and has good natural growth state in the field. As shown in fig. 1: the flower, leaf and fruit of passion flower of different varieties.

However, hybrid seeds of filial generations of the pinus dentata are easy to obtain, while the parent pinus dentata has certain resistance, but the flowers of the same plant are mutually hybridized and difficult to obtain seeds, the cutting seedling is difficult to graft due to hollow branches, and the root system of the cutting seedling is not as developed as the root system of the seedling.

In the prior art, in field planting, a planter usually adopts a mode of replacing seedlings once a year to deal with the occurrence of stem base rot. In practice, the seedlings of the yellow fruit passion fruit variety have slightly stronger stem base rot resistance than the seedlings of the purple fruit variety, and some companies or planters graft the purple fruit variety by using seedlings of yellow fruits as stocks, so that the stem base rot resistance is enhanced after grafting, the problems are relieved to a certain extent, but the disease onset time is delayed for about half a year. Therefore, how to obtain the resistant rootstock with good characters is still an important content in breeding research, and the important significance is achieved in cultivating the rootstock which has excellent resistance and is suitable for grafting.

The patent publication No. CN111248037A entitled "method for preventing and treating stem base rot of passion fruit" adopts the measures of removing disease spots, cleaning germs, applying medicine and the like to deal with the diseased passion fruit plants, and is a chemical agent prevention and treatment method;

the patent with the patent publication number of CN108353708B and the name of a method for preventing and treating the stem base rot of passion flower adopts physical and chemical medicament prevention and treatment methods such as disease part cutting, 75% alcohol cut disinfection, sterilization medicament smearing, recovery medicament smearing, white agent whitewashing and the like; aiming at the problem that passion fruit is susceptible to stem basal rot, compared with the two patents, the control thought is completely different.

Patent (publication No. CN201811584620.9) "a seedling cultivation method for enhancing the resistance of passion fruit plants to stem basal rot", which selects safflower passion flower (P.miniata) and tetraploid safflower passion flower with stronger resistance as rootstocks, and grafts different branches of high-quality passion fruit varieties as scions, thereby enhancing the capability of passion fruit seedlings to resist the stem basal rot, but the grafted seedlings using the rootstocks have weaker capability of sprouting the branches and more limited yield in field planting.

Aiming at the technical problems, the cultivation method of the passion fruit seedlings, which has the advantages of strong disease resistance of rootstocks, easiness in obtaining, strong stem and base rot resistance, luxuriant branches of the seedlings and high yield, is needed, so that the cost is reduced, and the yield is improved.

Disclosure of Invention

The invention aims to provide a cultivation method of passion fruit seedlings, in particular to a cultivation method of passion fruit seedlings which have strong stem rot resistance and luxuriant seedling branches.

The purpose of the invention is realized by the following technical scheme:

a cultivation method of passion fruit seedlings is characterized in that passion flower and passion flower with passion flower canna are used as parents, hybridization is conducted to obtain filial generation F1 which is used as a grafted stock, tender branches of passion fruit are used as scions, an oblique grafting method is adopted for grafting cultivation, and grafting survival is conducted to obtain the passion fruit seedlings capable of resisting stem rot.

Further, the cultivation method of the passion fruit seedlings comprises the following steps:

s1: selecting and pretreating rootstocks: selecting a filial generation F1 which has no plant diseases and insect pests, strong growth and branch thickness diameter of more than or equal to 0.2cm as a grafted stock, pruning the stock, keeping the height of the pruned stock above 15cm, and removing all branches and leaves above a pre-grafting point;

s2: pretreatment of scions: selecting tender passion fruit branches which are robust in growth, free of diseases and insect pests, strong in vitality and full in bud bodies and have the diameter of 0.2-0.5cm, cutting the branches into scions with leaf buds and the lengths of 8-15 cm by using a pruning shear, wherein the thickness of each scion is close to that of a stock, and at least one node is reserved on each scion so as to grow new buds from the nodes; leaves of the scions are kept half-size leaves to prevent excessive evaporation of water;

s3: grafting by oblique grafting: cutting 1/3-1/2 with thick branches at the lower end of the scion and the upper end of the rootstock respectively by using a grafting knife to form a smooth inclined plane with the length of 1-4 cm; the cutting surfaces of the stock and the scion are aligned, the cutting surfaces of the scion are tightly attached to the cutting surfaces of the stock by using the grafting clamp, the stock is clamped and fixed firmly, and meanwhile, the self-sealing bag is sleeved on the scion, so that on one hand, moisture is prevented from entering the wound during watering, infection and grafting failure are avoided, on the other hand, certain humidity can be kept for the leaves, and survival is guaranteed.

S4: and (3) management after grafting: watering is not needed within 2-3 days after grafting, and the wound infection caused by internal water seepage at the wound is prevented; timely watering is needed, and the buds germinated on the rootstocks are removed in time, so that nutrition is supplied to the scions in a centralized manner; after 7-15 days of grafting, the valve bag can be removed after the wound is healed; and (3) removing the grafting clips 1-2 months after grafting to prevent the grafting clips from influencing the growth of branches, and obtaining the passion fruit seedlings with the stem rot resistance after the grafting survives.

Further, the cut scions in S2 are placed in a sealed bag with humidity more than or equal to 80% for moisture preservation.

Further, the length of the valve bag in the S3 is larger than that of the scion, so that the wound cannot enter water when watering.

Furthermore, in the step S4, after grafting, management is performed, and the grafted plant is placed in a cultivation environment with humidity of about 80% to prevent graft wilting and graft survival from being affected.

Further, the diameter of the grafting clip in the step S3 should be slightly smaller than that of the wound junction, so that the clip is firmly fixed, and the scion is tightly attached to the rootstock.

Further, the rootstock in S1 is obtained by hybridizing passion flower with passion flower of green fruit, pollinating by adopting a conventional pollination mode, growing to obtain fruits, and cultivating the fruits by using seeds in the fruits. The filial generation F1 includes filial generation of the serrate green fruit (P.serrulata X P.maliformis) and filial generation of the serrate green fruit (P.maliformis X P.serrulata).

Further, the cultivation step of the rootstock filial generation F1 in the S1 comprises the following steps:

(1) selection and pretreatment of female parent: selecting healthy and disease-free buds of passion flower to be opened, opening the petals with tweezers, removing all pollen, firmly covering with waterproof kraft paper bags, and writing labels; bagging is to select a watertight kraft paper bag to prevent the pollinated pollen from being washed away by rain and prevent microminiature insects from entering with other pollen;

(2) selecting and pretreating male parents: selecting normal, healthy and disease-free flowers of passion fruit on the same day, and sweeping mature pollen into a clean disposable culture dish by using a clean writing brush;

(3) pollination: selecting windless morning, 8-10 o' clock, before the stigma secretions are not dried, smearing the passion flower pollen in the disposable culture dish in the step (2) on the stigma of the passion flower treated in the step (1) by using a clean writing brush, and bagging; the green fruit passion flower used for pollination is pollen collected on the same day; during pollination, windless weather is selected to prevent male parent confusion caused by the spread of other pollen by wind medium. And (3) management after pollination: 3-5 days after pollination, taking off a kraft paper bag, observing whether the ovary is expanded, preliminarily judging whether the pollination is successful, and replacing a nylon mesh bag with 30 x 20cm for flowers which are successfully pollinated so as to facilitate later observation;

(4) and (3) management after pollination: 3-5 days after pollination, taking off the kraft paper bag, observing whether the ovary is enlarged, and primarily judging

If the pollination is successful, replacing the flowers which are successfully pollinated with a nylon mesh bag of 30 x 20cm, so as to be convenient for later observation; the size of the passion flower fruits is 2-4cm, and the width of the nylon mesh bag is larger than the diameter of the fruits so as to prevent the fruits from growing;

(5) fruit harvesting and hybrid breeding: after pollination, fruits ripen and fall off about 60 days, the peel turns orange from green, and after the fruits fall off, the green serrate fruit hybrid seeds are obtained;

the green fruit-passionflower is used as a female parent and the passionflower denticulata is used as a male parent, and the green fruit-passionflower denticulata hybrid seeds are obtained by adopting the same way.

Further, the seedling cultivation of the hybrid seeds adopts a conventional seedling cultivation mode, and is carried out according to the following steps:

(1) pretreatment of the substrate: selecting a clean matrix, putting the matrix in a seeding pot with the thickness of about 3-5cm, and watering to completely wet the matrix; the used matrix only needs to be clean, the type is not limited, and the matrix can be turfy soil, vermiculite or mixed matrix in any proportion;

(2) seed cleaning: collecting green fruit pinus dentatus hybrid seeds or pinus dentatus green fruit hybrid seeds in fruits, and cleaning all pulp tissues surrounding the seeds;

(3) sowing and managing: uniformly scattering the pretreated seeds in a sowing basin, and covering a layer of wetted turfy soil with the thickness of about 0.5cm on the surface; after sowing, placing the seeds in a greenhouse with a shading net to avoid sunshine insolation, and watering the seeds once every 2 days;

(4) seedling management: after seeding for about 20 days, the cotyledon of the seed is unearthed, and when 3-4 true leaves grow out from the hybrid seedling, the seedling is transplanted to be a single plant; transplanting the seedlings to a greenhouse with sufficient sunlight after transplanting the seedlings for one week, and watering the seedlings once every 3 days; the seedlings which just move into the pot are placed under a shading net of a greenhouse, so that the seedlings are protected from being exposed to sunlight and are given a certain seedling revival period.

The cultivation of the seeds and seedlings of the filial generation of the green cherry filial generation adopts the same method as the filial generation of the green cherry filial generation, and only the male parent and the female parent are exchanged.

The invention has the beneficial effects that:

(1) the method creatively selects a filial generation F1 obtained by hybridizing passion flower with fine teeth and passion flower with green fruit with stronger resistance as a stock, selects tender branches of passion fruit with leaf buds as scions, adopts an oblique grafting method for grafting, and grafts the seedlings into live seedlings as passion fruit seedlings. The seedlings cultivated by the method can improve the capability of the passion fruit seedlings to resist the stem basal rot, not only utilizes the hybridization advantage that the posterity of interspecific hybridization is vigorous, but also integrates the excellent character of the stem basal rot resistance of the stock and the excellent fruit character of the scion, overcomes the defect of weak resistance of the passion fruit seedlings, reduces the cultivation and management cost in the passion fruit planting process, and ensures that a grower does not need to replace the seedlings once a year; the harvest time of the passion fruit is prolonged, the yield and the total economic value of the passion fruit are increased, and the method has important significance for promoting the large-scale planting of the passion fruit.

(2) The seedling cultivation method provided by the invention is simple to operate, low in cost, suitable for agricultural mass production, wide in market prospect and commercial value, and has important revelation for high-quality seedling cultivation of passionflower other plants.

(3) The stock that this patent adopted obtains easily through conventional interspecific hybridization, and prior art's stock relatively, the stock that this patent adopted more has the affinity strong, and the stock sprouts less and the scion sprouts the fast good characteristics of sprout after the grafting, and the sprout on the scion sprouts fast promptly, sprouts the reinforce, grafting survival rate is fast, the short advantage of the required time of becoming young. The inherent reason that the stock sprouts less is that the speed of the conduction tissue communication and healing between the stock and the scion is higher and better, and nutrient substances such as moisture absorbed by the root of the stock can be timely conveyed and transmitted to the related tissue part of the scion to promote the scion to grow new leaves, so that most nutrient substances are not accumulated in the tissues of the stock, and the stock is promoted to sprout more.

Drawings and description of the drawings:

FIG. 1 is a diagram of the flowers, leaves and immature fruits of different varieties of passion flower, wherein a, e and l in FIG. 1 are respectively a diagram of flowers, leaves and immature fruits of passion flower (P.serrulata); the symbols b, f and m in FIG. 1 are the flower, leaf and immature fruit of the filial generation of the green fruit with serration (P.maliformis X P.serrulata); the symbols c, g and n in FIG. 1 are the flower, leaf and immature fruit of the filial generation of the Eleocharis serrata x P.maliformis respectively; labeled d, h, o: maps of flowers, leaves, immature fruits of passion fruit (p. maliformis), respectively; in FIG. 1, i and k are the growth states of green fruit pint filial generation (P.maliformis X P.serrulata) seedlings with age of 7 months and 3 months of field planting, and the seedlings can grow to 2m high and 3m wide plants in crown width within half a year; the growth potential is very strong; reference numeral j in FIG. 1 is a comparison of mature fruits of passion flower (P.serrulata), passion fruit (P.maliformis), and filial generations of passion fruit (P.serrulata x P.maliformis), with passion flower on fruit on the left, filial generations of passion fruit on the middle, and passion flower on fruit on the right.

FIG. 2 is a comparison of the rootstock resistance experiment, in which the disease resistance is verified by inoculating Passiflora incarnata (P.serrulata) living body marked as a in FIG. 2, three strains on the left side in a are taken as a control, and three strains on the right side are taken as inoculated seedlings; in FIG. 2, b is a filial generation of green fruit serrate (P.maliformis X P.serrulata) in vivo inoculated to verify its disease resistance, the two strains on the left are Passiflora incarnata control seedlings, the two strains in the middle are Passiflora incarnata inoculated seedlings, and the two strains on the right are filial generation inoculated seedlings of green fruit serrate; FIG. 2 is a graph of stem of a P.serrulata X P.maliformis test seedling designated as c, the two strains on the left are controls, and the two strains on the right are inoculants; FIG. 2 d shows the filial generation (P.serrulata X P.maliformis) of the green dentate fruit, which is inoculated with the bacterium to verify the disease resistance, the two strains on the left are the control seedlings of Passiflora incarnata, the two strains in the middle are the inoculated seedlings of Passiflora incarnata, and the two strains on the right are the filial generation seedlings of the green dentate fruit; FIG. 2 is a graph of the stem of a P.serrata test seedling, in which the left 2 lines are the stem of a Passiflora incarnata inoculation seedling, the middle 2 lines are the stem of a Passiflora incarnata inoculation seedling, and the right 2 lines are the stem of a Passiflora incarnata control seedling; fig. 2 is a graph f showing the stem of the seedling of the green fruit serrate hybrid, which is represented by the two control seedlings of passion fruit, two inoculation seedlings of the green fruit serrate hybrid, and two control seedlings of the green fruit serrate hybrid from left to right.

Fig. 3 is a graph comparing the growth of seedlings of passion fruit cultured on different stocks, wherein fig. 3 is marked as a graph showing the seedling sprouting ability of the filial generation of the serrate green fruit as the stock, and fig. 3 is marked as B showing the seedling sprouting ability of the passion flower as the stock.

The specific implementation mode is as follows:

the present invention will be further described with reference to examples in order to facilitate understanding by those skilled in the art.

The examples are only illustrative and not restrictive, and the steps not specifically described in the examples are prior art and will not be described in detail herein.

Example 1 a method for cultivating a passion fruit rootstock,

firstly, cultivating filial generation of rootstock pinus green fruit, and pollinating by adopting a conventional pollination mode, wherein the method comprises the following steps:

(1) selection and pretreatment of female parent: selecting 10 healthy and disease-free passion flower buds to be opened in 2019, 9 and 20 days, opening the petals with tweezers, removing all pollen, firmly covering with a waterproof kraft paper bag, and writing a label;

(2) selecting and pretreating male parents: selecting normal open at 8-10 am of 21 am of 9/2019,

Cleaning mature pollen of healthy and disease-free flowers of passion fruit in a clean disposable culture dish by using a clean writing brush;

(3) pollination: applying pollen of passion fruit green onto stigma of passion fruit tree treated in 20 days in 9 months by using a clean writing brush 8-10 points earlier in 21 days in 2019 and 9 months, and bagging;

(4) and (3) management after pollination: and (4) taking off the kraft paper bag in 2019, 9 and 25 months, wherein the ovary of 8 pollinated passion flower buds is enlarged, and a nylon mesh bag with 30 x 20cm is replaced for the 8 flowers which are successfully pollinated.

(5) Harvesting fruits: collecting 1 mature and fallen hybrid fruits (P.serrulata x P.maliformis) in 11.5.5.5.5.2019, and taking out the fruits from a nylon mesh bag;

(6) pretreatment of the substrate: on the day of sowing, the substrate is subpackaged in a clean sowing basin with the thickness of about 3-5cm, and is watered to be completely wet;

(7) seed cleaning: breaking the pericarp in the step (5) by using a blade, taking out the seeds inside, and cleaning all pulp tissues wrapping the seeds to obtain 86 seeds in total;

(8) sowing and managing: uniformly spreading seeds in a sowing basin in the afternoon of 11 months and 5 days in 2019, and covering a layer of wetted turfy soil with the thickness of about 0.5cm on the surface; after sowing, placing the seeds in a greenhouse with a shading net to avoid sunshine insolation, and watering the seeds once every 2 days;

(9) seedling management: starting from 25 days 11 and 25 months in 2019, the cotyledons of the seeds are continuously unearthed, when 3-4 true leaves grow out from the hybrid seedlings, the seedlings are transplanted to be single plants, and 65 single plants are obtained in total by 18 days 1 and 18 months in 2020; after transplanting seedlings for one week, the seedlings are moved to a greenhouse with sufficient sunlight and watered once every 3 days.

2. Cultivation method of passion fruit seedlings

(1) Selection and pretreatment of rootstocks: selecting 30 pindus indica hybrid seeds with stem thickness of 0.2-0.3cm as the stock in 8 days 5 months in 2020, pruning the stock, and cutting off all branches and leaves with the stem thickness of more than 0.2cm to ensure that the stem height after pruning is 15-25 cm.

(2) Selection and pretreatment of scions: selecting tender branches of gypsophila paniculata which are strong in growth and free of diseases and insect pests as scions, cutting the tender branches with leaf buds by using a branch shear, cutting the tender branches into scions with the length of 10-15cm and the diameter of 0.2-0.3cm, ensuring that each scion has 1-2 leaf buds, cutting off 2/3 from leaves on the scion, placing the scions in self-sealing bags which are sprayed with moisture in advance, and moisturizing the scions.

(3) Grafting: and (3) cutting off 1/3-1/2 with thick branches from the lower end of the scion and the upper end of the rootstock respectively by using a grafting knife to form a smooth inclined plane with the length of 3-5 cm. The method comprises the steps of selecting scions and stocks with similar diameters, enabling the stocks and the sections of the scions to be tightly attached together by using a grafting clamp, and meanwhile, covering a No. 7 self-sealing bag on the scions to prevent external moisture from entering wounds and enable leaves to keep certain humidity, so that survival is guaranteed.

(4) And (3) management after grafting: after the grafting operation is finished, the grafted seedlings are placed under a sunshade net for management, watering is avoided as much as possible, and wound infection caused by internal moisture at the wound is prevented; and starting from 12 days 5 and 12 months in 2020, watering at proper time to prevent plant wilting caused by water shortage. Removing the self-sealing bag to prevent the limitation of the growth of the scion in 23 days of 5 months in 2020; and 7, 7 and 15 days in 2020, the grafting clips are removed to prevent the grafting clips from influencing the transverse growth of the branches. Within two months of grafting, timely observing the buds germinated by the removed rootstocks so as to supply nutrition to the scions in a centralized manner; by 20 days 7 and 20 months in 2020, the grafted seedlings total to 26 survivors.

Example 2 cultivation method of Passion fruit seedlings

The method comprises the following steps of firstly, cultivating filial generations of rootstock green fruit pinous teeth, and pollinating by adopting a conventional pollination mode:

(1) selection and pretreatment of female parent: selecting 15 healthy disease-free passion fruit buds to be opened in 2019, 10 months and 8 days, opening the petals with tweezers, removing all pollen, firmly covering with a waterproof kraft paper bag, and writing a label;

(2) selecting and pretreating male parents: selecting normal open healthy disease-free passion flower at 8-10 am in 2019, 10 and 9 months, and sweeping mature pollen into a clean disposable culture dish by using a clean writing brush;

(3) pollination: applying pollen of passion fruit green onto stigma of passion fruit which is treated in 10 month and 2 days by using a clean writing brush at 8-10 o' clock in the morning of 10 month and 9 days in 2019, and bagging;

(4) and (3) management after pollination: taking down the kraft paper bag in 2019, 9, 25 and 15 pollinated passion fruit, wherein the ovary of each 15 pollinated flowers is enlarged, and a nylon mesh bag of 30 x 20cm is replaced for the 15 flowers which are successfully pollinated;

(5) harvesting fruits: collecting 1 mature and fallen hybrid fruits (P.maliformis X P.serrulata) in the morning of 9 months in 2020, and taking out the fruits from a nylon mesh bag;

(6) pretreatment of the substrate: on the day of sowing, the substrate is subpackaged in a clean sowing basin with the thickness of about 3-5cm, and is watered to be completely wet;

(7) seed cleaning: breaking the peel in the step (5) by using a blade, taking out the seeds inside, and cleaning all pulp tissues wrapping the seeds to obtain 375 seeds in total;

(8) sowing and managing: uniformly spreading the seeds in a sowing basin in the afternoon of the same day, and covering a layer of wet turfy soil with the thickness of about 0.5cm on the surface; after sowing, placing the seeds in a greenhouse with a shading net to avoid sunshine insolation, and watering the seeds once every 2 days;

(9) seedling management: the cotyledon of the seed begins to come out continuously in 6 days 4 and 6 months 2020, when 3-4 true leaves grow out from the hybrid seedling, the seedling is transplanted to a single plant, and 302 single plants are obtained in total by 5 and 25 days 5 and 25 months 2020; after transplanting seedlings for one week, the seedlings are moved to a greenhouse with sufficient sunlight and watered once every 3 days.

Second, cultivation method of passion fruit seedlings

(1) Selection and pretreatment of rootstocks: selecting 30 green fruit serrate hybrids with the stem thickness of 0.2-0.3cm as the stock in 5-15 months in 2020, pruning the stock, and cutting off all branches and leaves with the stem thickness of more than 0.2cm to ensure that the stem height after pruning is 15-25 cm.

(2) Selection and pretreatment of scions: the method comprises the steps of selecting robust and disease and insect pest free tender branches of golden fruits as scions, using a branch shear to cut the tender branches with leaf buds, then cutting the tender branches into scions with the length of 10-15cm and the diameter of 0.2-0.3cm, ensuring that each scion has 1-2 leaf buds, cutting off 2/3 from leaves on the scion, then placing the scions in self-sealing bags which are sprayed with moisture in advance, and preserving moisture of the scions.

(3) Grafting: and (3) cutting off 1/3-1/2 with thick branches from the lower end of the scion and the upper end of the rootstock respectively by using a grafting knife to form a smooth inclined plane with the length of 3-5 cm. The method comprises the steps of selecting scions and stocks with similar diameters, aligning the cut surfaces of the stocks and the scions, tightly attaching the cut surfaces of the stocks and the scions by using a grafting clamp, and simultaneously sleeving a No. 7 self-sealing bag on the scions to prevent external moisture from entering wounds and enable leaves to keep certain humidity, so that survival is guaranteed.

(4) And (3) management after grafting: after the grafting operation is finished, the grafted seedlings are placed under a sunshade net for management, watering is avoided as much as possible, and wound infection caused by internal moisture at the wound is prevented; and starting from 19/5/2020, watering at proper time to prevent plant wilting due to water shortage. Removing the self-sealing bag to prevent the limitation of the growth of the scion within 30 days of 5 months in 2020; and 7, 7 and 20 days in 2020, the grafting clips are removed to prevent the grafting clips from influencing the transverse growth of the branches. Within two months of grafting, timely observing the buds germinated by the removed rootstocks so as to supply nutrition to the scions in a centralized manner; by 25 days 7 and 25 months in 2020, the grafted seedlings totally survive 29 plants.

Example 3 comparison of rootstock resistance experiments

The resistance of the rootstocks is compared by comparing passion flower, filial generations of green fruit and green fruit, in-vivo inoculation experiments of filial generations of green fruit and the experimental seedling stem comparison map.

And (3) verifying the stem rot resistance of the filial generation of the green dentate fruits by adopting a living body inoculation method. Inoculating Fusarium solani (F.solani) to PDA culture medium, placing in a constant temperature incubator at 28 deg.C, culturing in reverse dark for 3-5 days, and beating the culture medium with colony to obtain small bacterial cake with diameter of 60mm with sterile perforator when the colony grows to half of the culture medium. Selecting 20 pindus green-fruit hybrid seeds with the seedling age of 4-6 months and the consistent growth potential, cutting a wound of 0.5cm multiplied by 0.2cm at the stem base part of the pindus green-fruit hybrid seeds by a blade, placing a small fungus cake with the front side facing the wound at the wound of 10 strains, and fixing the fungus cake at the wound by a sealing film; in addition, PDA culture medium which is not inoculated and has the diameter of 60mm is placed at the wound of 10 strains, and the culture medium is fixed at the wound by using a sealing film; in addition, 20 purple fruit passion fruit plants with the same seedling age as filial generations of the prunus serrata are selected as experimental controls, wherein 10 purple fruit passion fruit plants are inoculated with bacteria, and 10 purple fruit passion fruit plants are not inoculated with bacteria. All the experimental seedlings were cultured in a climatic chamber at 30 ℃ and 70% humidity to observe the onset of disease and the condition of the disease. The experiment was repeated three times.

FIG. 2 is a comparison of the rootstock resistance experiment, in which FIG. 2 shows a disease resistance test by inoculating Passiflora incarnata (P.serrulata) living body with three strains on the left as control and three strains on the right as inoculated vaccine. The experimental result shows that: the growth performance of the passion flower bud after 20 days of inoculation is good. FIG. 2 b shows the progeny of the green fruit serration cross (P. maliformis. times. P. serrulata) for which disease resistance was verified by live inoculation, the two left lines were the Passiflora incarnata control seedlings, the two middle lines were the Passiflora incarnata inoculation seedlings, and the two right lines were the progeny of the green fruit serration cross. The experimental result shows that: when the Passiflora incarnata inoculation seedlings die due to morbidity, the generation inoculation seedlings grow well after the green fruit serrate hybridization. FIG. 2 shows the stem of the seedling of the filial generation of the green dentate fruit (P.serrulata X P.maliformis) as reference, the left two plants are the passion fruit passionflower control, and the right two plants are the filial generation of the green dentate fruit. As a result, it was found that: the rhizosphere bacteria of the inoculated plants are rarely infected, and the resistance of the filial generation of the green dentate fruit to the stem rot is stronger than that of the passion fruit. FIG. 2 d shows the filial generation of the green dentate fruit (P.serrulata X P.maliformis) for in vivo inoculation to verify disease resistance, the two strains on the left are the passion fruit control seedlings, the two strains in the middle are the passion fruit inoculation seedlings, and the two strains on the right are the filial generation inoculation seedlings of the green dentate fruit. As a result, it was found that: when the passion fruit flower inoculation seedlings are completely withered, the generation inoculation seedlings after the green serrate fruit hybridization all grow well, and the disease resistance of the filial generation seedlings of the green serrate fruit hybridization is stronger than that of the passion fruit flower seedlings. FIG. 2 shows a stem map of a P.serrata test seedling, in which the left 2 lines are the stems of the Passiflora incarnata inoculated seedlings, the middle 2 lines are the stems of the Passiflora incarnata inoculated seedlings, and the right 2 lines are the stems of the Passiflora incarnata control seedlings. As a result, it was found that: when the bacterial plaque of the passion flower of purple fruit is infected at the whole stem base, the passion flower of dens is rarely infected, which shows that the passion flower of dens has certain capability of resisting stem base rot, and the disease resistance of the passion flower of dens is stronger than that of the passion flower of purple fruit. FIG. 2 shows a stem diagram of a P.maliformis X P.serrulata test seedling, which is represented by f, wherein two control seedlings of Passiflora incarnata, two inoculation seedlings of green fruit serrate hybrid, and two control seedlings of green fruit serrate hybrid are shown from left to right. As a result, it was found that: the stem base of the inoculated strain plant of the filial generation of the green fruit serrate is rarely infected by the bacteria, and the resistance of the filial generation of the green fruit serrate to stem rot is stronger than that of passion fruit. The experimental result shows that the filial generation of the green fruit serration has stronger capability of resisting the stem basal rot and can be used as a disease-resistant stock.

Example 4 influence of different rootstock seedling raising methods on the shoot initiation ability of passion fruit

The difference of the cultivation method lies in that the stocks are different, T1 takes the filial generation of the spikelet green fruit as the stock, takes the passion fruit branch as the scion, uses the grafting clip, adopts the oblique grafting method for grafting, the survival rate of the grafted seedling is high, the survival rate is fast, the growth state of the field planting is good, and the result is shown in figure 3A; t2 adopts passion flower as rootstock, the same method is used for grafting and planting, the growth state of the field planting is good, and the result is shown in figure 3B. Compared with T2, the survival time of T1 grafting is shortened to about 15 days from about 30 days, and the sprouts on the rootstock at the lower part of the grafting opening of the T1 rootstock after grafting are relatively less, so that the workload of removing the rootstock sprouts can be reduced. Therefore, the rootstock adopted by the method has the excellent characteristics of strong affinity, less rootstock germination and quick scion sprouting sprout after grafting, namely the advantages of quick sprouting speed, strong sprouting force, quick grafting survival speed and short seedling time. The internal reason that the stock sprouts less is that the speed of the conduction tissue communication and healing between the stock and the scion is higher and better, and nutrient substances such as moisture absorbed by the root of the stock can be timely conveyed and transmitted to the related tissue part of the scion to promote the scion to grow new leaves, so that most nutrient substances are unlikely to be accumulated in the tissues of the stock, more sprouts of the stock are promoted, and the workload of rejecting the stock sprouts is reduced. This patent does not need artificial intervention to make stock bud few when the grafting through the effective selection of stock, and the scion bud is fast and many, and this is very favorable to grafting seedling.

As can be seen from FIG. 3, the passion fruit cultivated by the method has a higher branching capability than that of a seedling cultivated by taking passion flower as a rootstock.

Claims (10)

1. A cultivation method of passion fruit seedlings is characterized by comprising the following steps: the method takes passion flower, passion flower and passion flower with serrate as parents, the filial generation F1 obtained by mutual hybridization is taken as a grafted stock, the tender branch of passion fruit is taken as a scion, the grafting cultivation is carried out by adopting an oblique grafting method, and the passion fruit seedling is obtained after the grafting survival.

2. The cultivation method of the passion fruit seedlings according to claim 1, characterized by comprising the following steps: the filial generation F1 comprises filial generation P.serrulata X P.maliformis and filial generation P.maliformis X P.serrulata.

3. The cultivation method of the passion fruit seedlings according to claim 2, characterized by comprising the following steps: the method comprises the following steps:

s1: selecting and pretreating rootstocks: selecting a filial generation F1 which has no plant diseases and insect pests, strong growth and branch thickness diameter of more than or equal to 0.2cm as a grafted stock, pruning the stock, keeping the height of the pruned stock above 15cm, and removing all branches and leaves above a pre-grafting point;

s2: pretreatment of scions: selecting tender passion fruit branches which are robust in growth, free of diseases and insect pests, strong in vitality and full in bud bodies and have the diameter of 0.2-0.5cm, cutting the branches into scions with leaf buds and the lengths of 8-15 cm by using a pruning shear, wherein the thickness of each scion is close to that of a stock, and at least one node is reserved on each scion so as to grow new buds from the nodes; leaves of the scions are kept half-size leaves;

s3: grafting by oblique grafting: cutting 1/3-1/2 with thick branches at the lower end of the scion and the upper end of the rootstock respectively by using a grafting knife to form a smooth inclined plane with the same length of 1-4 cm; aligning the stock with at least one side of the section of the scion, using a grafting clamp with a proper size to enable the scion cut surface to be tightly attached to the stock cut surface, simultaneously using a self-sealing bag to be sleeved on the scion for moisturizing, avoiding placing the scion in a full-sunlight environment after grafting is finished, and being suitable for placing the scion in a semi-shaded environment;

s4: and (3) management after grafting: watering is avoided as much as possible within 2-3 days after grafting, so that the wound is prevented from being infected by water seeping from the wound; timely watering is needed, and the buds germinated on the rootstocks are removed in time, so that nutrition is supplied to the scions in a centralized manner; after 7-15 days of grafting, the valve bag can be removed after the wound is healed; and (4) removing the grafting clip 1-2 months after grafting, and obtaining the passion fruit seedlings after grafting survival.

4. The cultivation method of the passion fruit seedlings according to claim 3, characterized by comprising the following steps: and (3) placing the pruned scions in the S2 in a sealed bag with the humidity of more than or equal to 80% for moisture preservation.

5. The cultivation method of the passion fruit seedlings according to claim 3, characterized by comprising the following steps: the length of the self-sealing bag in the S3 is larger than that of the scion.

6. The cultivation method of the passion fruit seedlings according to claim 3, characterized by comprising the following steps: and (3) performing management after grafting in S4, and placing the grafted plant in a cultivation environment with the humidity of about 80%.

7. The cultivation method of the passion fruit seedlings according to claim 3, characterized by comprising the following steps: the diameter of the grafting clip in the S3 is slightly smaller than that of the wound junction, so that the section of the scion is tightly attached to the section of the rootstock.

8. The cultivation method of the passion fruit seedlings according to claim 3, characterized by comprising the following steps: the rootstock filial generation F1 in the S1 is obtained by hybridizing passion flower with passion fruit, growing fruits after pollination is carried out by adopting a conventional pollination mode, and cultivating the fruits by using seeds in the fruits.

9. A cultivation method of passion fruit seedlings according to claim 8, characterized in that: the cultivation steps of the rootstock hybrid progeny seeds in the S1 are as follows:

(1) selection and pretreatment of female parent: selecting healthy and disease-free buds of passion flower to be opened, opening the petals with tweezers, removing all pollen, firmly covering with waterproof kraft paper bags, and writing labels;

(2) selecting and pretreating male parents: selecting normal, healthy and disease-free flowers of passion fruit on the same day, and sweeping mature pollen into a clean disposable culture dish by using a clean writing brush;

(3) pollination: selecting windless morning, 8-10 o' clock, before the stigma secretions are not dried, smearing the passion flower pollen in the disposable culture dish in the step (2) on the stigma of the passion flower treated in the step (1) by using a clean writing brush, and bagging; the green fruit passion flower used for pollination is pollen collected on the same day; selecting windless weather during pollination to prevent male parent confusion caused by other pollen propagating by wind medium;

(4) and (3) management after pollination: 3-5 days after pollination, taking off a kraft paper bag, observing whether the ovary is expanded, preliminarily judging whether the pollination is successful, and replacing a nylon mesh bag with 30 x 20cm for flowers which are successfully pollinated so as to facilitate later observation; the size of the passion flower fruits is 2-4cm, and the width of the nylon mesh bag is larger than the diameter of the fruits so as to prevent the fruits from growing;

(5) fruit harvesting and hybrid breeding: after pollination for about 60 days, the fruits mature and fall off, the peel turns to orange from green, after the fruits fall off, the fruits in the bag are collected to obtain the green serrate fruit hybrid seeds, and the obtained fruits are used for carrying out seedling cultivation of the green serrate fruit hybrid seeds;

the green fruit-passionflower is used as a female parent and the passionflower denticulata is used as a male parent, and the green fruit-passionflower denticulata hybrid seeds are obtained by adopting the same way.

10. A cultivation method of passion fruit seedlings according to claim 9, characterized by comprising the following steps: the seedling cultivation of the stock hybrid F1 adopts a conventional seedling cultivation mode to carry out seedling cultivation, and comprises the following steps:

(1) pretreatment of the substrate: selecting a clean matrix, putting the matrix in a seeding pot with the thickness of about 3-5cm, and watering to completely wet the matrix; the used matrix only needs to be clean, the type is not limited, and the matrix can be turfy soil, vermiculite or mixed matrix in any proportion;

(2) seed cleaning: collecting the green cherry hybrid seeds or green cherry hybrid seeds in the fruits, and cleaning all pulp tissues surrounding the seeds;

(3) sowing and managing: uniformly scattering the pretreated seeds in a sowing basin, and covering a layer of wetted turfy soil with the thickness of about 0.5cm on the surface; after sowing, placing the seeds in a greenhouse with a shading net to avoid sunshine insolation, and watering the seeds once every 2 days;

(4) seedling management: after seeding for about 20 days, the cotyledon of the seed is unearthed, and when 3-4 true leaves grow out from the hybrid seedling, the seedling is transplanted to be a single plant; transplanting the seedlings to a greenhouse with sufficient sunlight after transplanting the seedlings for one week, and watering the seedlings once every 3 days; the seedlings which just move into the pot are placed under a shading net of a greenhouse, so that the seedlings are protected from being exposed to sunlight and are given a certain seedling revival period.

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