Method for high yield of safflower shaddock

Abstract

This invention primarily relates to a method for increasing the yield of red-flowered wrinkled papaya. A method for increasing the yield of red-flowered wrinkled papaya has been invented. Based on the characteristics of red-flowered wrinkled papaya, which has a low natural fruit set rate and high physiological fruit drop, two artificial pollination methods are used: flower thinning and pollination wiping, and spraying with pollen. The water spraying technique used in Hubei crabapple for pollination and growth promotion can significantly increase the fruit set rate and yield of red-flowered wrinkled papaya, increasing the yield per plant from 150.7 g to over 5017.91 g. It also maintains the original varietal characteristics and quality performance, solving the problem of low yield in red-flowered wrinkled papaya.

Description

Technical Field

[0001] This invention belongs to the field of plant production technology, and in particular relates to a method for increasing the yield of safflower wrinkled papaya. Background Technology

[0002] According to the Chinese Pharmacopoeia, papaya originates from the dried, nearly mature fruit of *Chaenomeles speciosa* (Sweet) Nakai, a plant in the Rosaceae family. Currently, the main medicinal papaya producing areas in China are Changyang in Hubei Province, Xuancheng in Anhui Province, and Qijiang in Chongqing Province, where the papayas are commonly known as Ziqiu papaya, Xuan papaya, and Sichuan papaya, respectively. Over long-term cultivation, different varieties of papaya have emerged. The three major producing areas of Changyang in Hubei, Xuancheng in Anhui, and Qijiang in Chongqing all have two main groups: pink-flowered and bright red-flowered. A noteworthy phenomenon is that all three producing areas refer to the bright red-flowered group as "medicinal papaya" or simply "medicinal melon," believing its medicinal value to be higher than that of the pink-flowered group. However, the bright red-flowered group is currently endangered and requires stronger protection. The main reason for its endangerment is that this group produces almost no fruit, leading to a gradual decline in its cultivation and a sharp decrease in its numbers. Therefore, the best way to protect it is to ensure its high yield.

[0003] The characteristics and quality of Ziqiu papaya are significantly different from those of common wrinkled-skin papaya. Therefore, some scholars believe that Ziqiu papaya is a variant of wrinkled-skin papaya or a natural hybrid of wrinkled-skin papaya and hairy-leaved papaya. The most significant characteristic is that the vast majority of ovules in Ziqiu papaya fail to develop normally, resulting in virtually no seeds in the fruit. Over long-term cultivation, different types of Ziqiu papaya have differentiated. The common type has pink flowers, oblong-cylindrical fruit, almost no formed seeds, and occasionally 1-2 seeds. Another type may have bright red flowers; the fruit is sub-round or cylindrical, and compared to the pink-flowered wrinkled-skin papaya of the same period, the fruit is yellowish-green with prominent warts, almost no formed seeds, and occasionally 1-2 seeds.

[0004] The low natural fruit set rate of red-flowered wrinkled papaya is mainly due to two reasons: low pollination rate and high physiological fruit drop rate. This invention specifically addresses these problems. Summary of the Invention

[0005] In response to the problems of low pollination rate and high physiological fruit drop rate of red-flowered wrinkled papaya, the present invention provides a method for increasing the yield of red-flowered wrinkled papaya. The technical solution is as follows: (1) Planting: Select fertile, moist and well-drained land as planting field and level the land; after leveling the land, dig several planting holes, apply 10-20 kg of base fertilizer and soil mixture to each planting hole, and transplant the papaya seedlings into the planting holes;

[0006] Preferably, different red-flowered wrinkled papaya trees of the same age and size were selected from the plantation for the experiment;

[0007] (2) Powdering: At the beginning of the flowering period of the red-flowered wrinkled papaya, thin out the flowers and use the thinned flowers to directly and lightly rub the open flowers so that the pollen is applied to the pistil of the open flowers.

[0008] (3) Spraying: Collect pollen from the thinned flowers, add water extract of Malus hupehensis, and spray it on the flowers once during the initial flowering period and once during the full flowering period.

[0009] (4) Fruit preservation: Spray the water extract of Malus hupehensis onto the fruit during the initial fruiting period.

[0010] The planting hole spacing in step (1) is (1.8~2.2m)×(1.4~1.6m), and the depth is 60~70cm.

[0011] The base fertilizer mentioned in step (1) is one of the farmyard manures mainly composed of well-rotted pen, fence, compost, and biogas residue. It consists of a mixture of 2000-3000 kg of livestock manure, bedding straw and feed residue per mu, plus 20-30 kg of superphosphate and 5-10 kg of potassium sulfate (or 30-50 kg of wood ash).

[0012] The thinning of flowers mentioned in step (2) means retaining 3 to 5 flowers in the middle of each inflorescence.

[0013] The preparation method of the water extract of Malus hupehensis described in steps (3) and (4) is as follows: Malus hupehensis is used as raw material, and it is extracted by heating in a water bath and concentrated to a phlorizin content of 2 wt%. When using it, the phlorizin content is adjusted to 0.1-1.0 wt%.

[0014] Pest and disease control, fertilizer and water management, etc., are carried out according to routine procedures. Harvest when the outer skin of the red-flowered wrinkled papaya turns greenish-yellow. Calculate the yield from 5 representative trees (what traits are considered representative). Take three fruits from each of the 5 trees, cut them in half, dry them at 60℃ until the moisture content is about 80%, slice them, and continue drying until the moisture content is less than 10%. Determine the content of protocatechuic acid, chlorogenic acid, oleanolic acid, and ursolic acid.

[0015] The beneficial effects of this invention are:

[0016] (1) Characteristic components in the leaves of *Malus hupehensis* include phlorizin, 3-hydroxyphlorizin, and phloretin. The measured values ​​were phlorizin 3.97%, 3-hydroxyphlorizin 0.26%, and phloretin 0.33%. Aqueous extracts of *Malus hupehensis* with phlorizin content of 0.1–1% can improve fruit setting and retention in *Chaenomeles speciosa*, thus promoting high yield.

[0017] (2) Both the untreated and the red-flowered wrinkled papaya obtained by different treatments had good quality. The total amount of oleanolic acid and ursolic acid far exceeded the standard of 0.5% in the Chinese Pharmacopoeia and was higher than that of pink and white-flowered wrinkled papaya, indicating that the treatment method can ensure that the quality of the fruit does not decline while achieving high yield.

[0018] (3) Both powdering and spraying can improve the fruit setting of red-flowered wrinkled papaya and promote high yield. Spraying with a water extract of *Malus hupehensis* containing 0.1-1% phlorizin can further improve the fruit setting ability of red-flowered wrinkled papaya. Compared with simple powdering, the yield per plant increased from 1959.54g to 4736.27g, an increase of 141.7%.

[0019] (4) Spraying the fruit with the water extract of *Malus hupehensis* on the *Papaver somniferum* 'Red Flower Wrinkled Skin' can protect the fruit and promote high yield. The yield per plant increased from 4211.85g to 5017.91g, an increase of 19.14%. Attached Figure Description

[0020] Figure 1 This is an HPLC chromatogram of the water extract of Malus hupehensis leaves. Detailed Implementation

[0021] The present invention will be further described in detail below with reference to specific embodiments.

[0022] Example 1

[0023] Determination of active substance content in *Malus hupehensis*

[0024] 1.1 Experimental Materials

[0025] The young leaves of Malus hupehensis (Pamp.) Rehd. were provided by Enshi Selenium-containing Malus Biotechnology Co., Ltd. They were identified by Professor Wang Yubing of Three Gorges University as belonging to Malus hupehensis.

[0026] 1.2 Reagents

[0027] Phlorizin, 3-hydroxyphlorizin, and phloretin (purity ≥98%) were provided by the Hubei Provincial Key Laboratory of Natural Product Research and Utilization (Three Gorges University). Protocatechuic acid (purity ≥98.0%) was provided by Chengdu Professional Biotechnology Co., Ltd. Chlorogenic acid reference standard (purity ≥98.0%) was provided by the National Institutes for Food and Drug Control. Chromatographic methanol and acetonitrile were provided by Thermo Fisher Scientific, USA. Phosphoric acid was provided by Tianjin Kemei Chemical Reagent Co., Ltd. All other reagents were of analytical grade.

[0028] 1.3 Experimental Apparatus

[0029] See Table 1.

[0030] Table 1 Main Instruments and Equipment

[0031]

[0032] 2 Experimental Methods and Results

[0033] 2.1 Chromatographic conditions for constructing the standard curve: Column: 5C18-MS-II column (4.6ID×250mm), Cosmed, Japan; Mobile phase: acetonitrile (A)-water (B) gradient elution, A increased from 10% to 100% from 0-25min, and decreased from 100% to 10% from 25min-30min; Detection wavelength: 287nm; Column temperature: 28℃; Injection volume: 10μL; Flow rate: 1.0ml / min.

[0034] Preparation of reference solutions: Accurately weigh 100 mg each of phlorizin, 3-hydroxyphlorizin, and phloretin, dissolve them thoroughly in methanol, and then dilute to 100 mL in a volumetric flask. Shake well to obtain a reference solution with a mass concentration of 1.00 mg / mL. Prepare a series of reference solutions with mass concentrations of 25, 50, 100, 200, 300, 400, and 500 μg / mL using methanol.

[0035] Standard curve: 10 μL of the standard solution was injected and analyzed under experimental chromatographic conditions. Three injections were performed, and the average peak area was calculated. Linear regression was conducted with the mass concentrations (X) of phlorizin, 3-hydroxyphlorizin, and phloretin as the abscissa and the peak area (Y) as the ordinate to establish regression equations (n ​​= 5). Phlorizin regression equation: Y = 0.2915X - 0.2598, R² = 0.9998, linear range: 25–500 μg / mL; 3-hydroxyphlorizin regression equation: Y = 0.112X + 1.2641, R² = 0.9991, linear range: 25–500 μg / mL; Phloretin regression equation: Y = 0.4592X - 1.325, R² = 0.9995, linear range: 25–500 μg / mL.

[0036] 2.2 Determination of Active Substance Content in Malus hupehensis

[0037] Accurately weigh 1g of *Malus hupehensis* leaves, soak in 20ml of chromatographic methanol for 30min, and ultrasonically extract at room temperature for 30min (power 400W, frequency 40kHz). Let stand, collect the supernatant, filter through a 0.45μm microporous membrane, and collect the filtrate to obtain the *Malus hupehensis* test solution. Determine the contents of phlorizin, 3-hydroxyphlorizin, and phloretin under experimental chromatographic conditions, and calculate the contents based on the linear regression equation. The results showed that phlorizin in *Malus hupehensis* leaves was 3.97%, 3-hydroxyphlorizin was 0.26%, and phloretin was 0.33%.

[0038] Example 2

[0039] Preparation of water extract of Malus hupehensis

[0040] Weigh 1 kg of *Malus hupehensis*, add 10 kg of water, and extract twice by heating in a 90℃ water bath. Each time, after reaching 90℃, maintain the temperature for 30 minutes. Filter and combine the two filtrates. Concentrate to a phlorizin content of 2 wt%, and store at 4℃ for later use.

[0041] Example 3

[0042] (1) Planting: Select fertile, moist and well-drained land as planting fields and level the land; after leveling the land, plant 110 plants per mu, apply 10-20 kg of base fertilizer and soil mixture to each planting hole, and transplant the papaya seedlings into the planting holes.

[0043] Furthermore, 30 different red-flowered wrinkled papaya trees of the same age and size were selected from the plantation for an experiment;

[0044] (2) Powdering: At the beginning of the flowering period of the red-flowered wrinkled papaya, thin out the flowers and use the thinned flowers to directly and lightly rub the open flowers so that the pollen is applied to the pistil of the open flowers.

[0045] (3) Spraying: Wash 500g of the thinned flowers with 20kg of water to collect pollen (about 20g), remove petals and other impurities, add the water extract of Malus hupehensis prepared in Example 2, adjust the phlorizin content to 0.1%, spray it on the flowers once at the initial flowering stage and once at the full flowering stage, with each spray volume being 0.5L.

[0046] The planting hole spacing in step (1) is (1.8~2.2m)×(1.4~1.6m), and the depth is 60~70cm.

[0047] The base fertilizer mentioned in step (1) is well-rotted manure, a mixture of 2000-3000 kg of livestock manure, bedding straw and feed residue per mu, plus 20-30 kg of superphosphate and 5-10 kg of potassium sulfate (or 30-50 kg of wood ash).

[0048] The thinning of flowers mentioned in step (2) means retaining 3 to 5 flowers in the middle of each inflorescence.

[0049] Pest and disease control, fertilizer and water management, etc. are carried out in accordance with the usual procedures. Harvest when the skin of the red-flowered wrinkled papaya turns greenish-yellow.

[0050] Example 4

[0051] The experimental procedure was the same as in Example 3, except that "the aqueous extract of Malus hupehensis prepared in Example 2 was adjusted to have a phlorizin content of 0.1%" was changed to "the aqueous extract of Malus hupehensis prepared in Example 2 was adjusted to have a phlorizin content of 0.5%".

[0052] Example 5

[0053] The experimental procedure was the same as in Example 3, except that "the aqueous extract of Malus hupehensis prepared in Example 2 was adjusted to have a phlorizin content of 0.1%" was changed to "the aqueous extract of Malus hupehensis prepared in Example 2 was adjusted to have a phlorizin content of 1.0%".

[0054] Example 6

[0055] (1) Planting: Select fertile, moist and well-drained land as planting fields and level the land; after leveling the land, plant 110 plants per mu, apply 10-20 kg of base fertilizer and soil mixture to each planting hole, and transplant the papaya seedlings into the planting holes.

[0056] Furthermore, 30 different red-flowered wrinkled papaya trees of the same age and size were selected from the plantation for an experiment;

[0057] (2) Powdering: At the beginning of the flowering period of the red-flowered wrinkled papaya, thin out the flowers and use the thinned flowers to directly and lightly rub the open flowers so that the pollen is applied to the pistil of the open flowers.

[0058] (3) Fruit preservation: Spray the fruit with a water extract of 0.5% phlorizin at the initial fruiting stage, with a spray volume of 1L per tree.

[0059] The planting hole spacing in step (1) is (1.8~2.2m)×(1.4~1.6m), and the depth is 60~70cm.

[0060] The base fertilizer mentioned in step (1) is well-rotted manure, a mixture of 2000-3000 kg of livestock manure, bedding straw and feed residue per mu, plus 20-30 kg of superphosphate and 5-10 kg of potassium sulfate (or 30-50 kg of wood ash).

[0061] The thinning of flowers mentioned in step (2) means retaining 3 to 5 flowers in the middle of each inflorescence.

[0062] Pest and disease control, fertilizer and water management, etc. are carried out in accordance with the usual procedures. Harvest when the skin of the red-flowered wrinkled papaya turns greenish-yellow.

[0063] Example 7

[0064] (1) Planting: Select fertile, moist and well-drained land as planting fields and level the land; after leveling the land, plant 110 plants per mu, apply 10-20 kg of base fertilizer and soil mixture to each planting hole, and transplant the papaya seedlings into the planting holes.

[0065] Furthermore, 30 different red-flowered wrinkled papaya trees of the same age and size were selected from the plantation for an experiment;

[0066] (2) Powdering: At the beginning of the flowering period of the red-flowered wrinkled papaya, thin out the flowers and use the thinned flowers to directly and lightly rub the open flowers so that the pollen is applied to the pistil of the open flowers.

[0067] (3) Spraying: Wash 500g of the thinned flowers with 20kg of water to collect pollen (about 20g), remove petals and other impurities, add the water extract of Malus hupehensis prepared in Example 2, adjust the phlorizin content to 0.5%, spray it on the flowers once at the initial flowering stage and once at the full flowering stage, with each spray volume being 0.5L;

[0068] (4) Fruit preservation: Spray the fruit with a water extract of 0.5% phlorizin at the initial fruiting stage, with a spray volume of 1L per tree.

[0069] The planting hole spacing in step (1) is (1.8~2.2m)×(1.4~1.6m), and the depth is 60~70cm.

[0070] The base fertilizer mentioned in step (1) is well-rotted manure, a mixture of 2000-3000 kg of livestock manure, bedding straw and feed residue per mu, plus 20-30 kg of superphosphate and 5-10 kg of potassium sulfate (or 30-50 kg of wood ash).

[0071] The thinning of flowers mentioned in step (2) means retaining 3 to 5 flowers in the middle of each inflorescence.

[0072] Pest and disease control, fertilizer and water management, etc. are carried out in accordance with the usual procedures. Harvest when the skin of the red-flowered wrinkled papaya turns greenish-yellow.

[0073] Example 8

[0074] (1) Planting: Select fertile, moist and well-drained land as planting fields and level the land; after leveling the land, plant 110 plants per mu, apply 10-20 kg of base fertilizer and soil mixture to each planting hole, and transplant the papaya seedlings into the planting holes.

[0075] Furthermore, 30 different red-flowered wrinkled papaya trees of the same age and size were selected from the plantation for an experiment;

[0076] (2) Flower thinning: Flower thinning should begin at the initial flowering stage of the red-flowered wrinkled quince;

[0077] (3) Spraying: Wash 500g of the thinned flowers with 20kg of water to collect pollen (about 20g), remove petals and other impurities, add the water extract of Malus hupehensis prepared in Example 2, adjust the phlorizin content to 0.5%, spray it on the flowers once at the initial flowering stage and once at the full flowering stage, with each spray volume being 0.5L;

[0078] (4) Fruit preservation: Spray the fruit with a water extract of 0.5% phlorizin at the initial fruiting stage, with a spray volume of 1L per tree.

[0079] The planting hole spacing in step (1) is (1.8~2.2m)×(1.4~1.6m), and the depth is 60~70cm.

[0080] The base fertilizer mentioned in step (1) is farmyard manure.

[0081] The thinning of flowers mentioned in step (2) means retaining 3 to 5 flowers in the middle of each inflorescence.

[0082] Pest and disease control, fertilizer and water management, etc. are carried out in accordance with the usual procedures. Harvest when the skin of the red-flowered wrinkled papaya turns greenish-yellow.

[0083] Comparative Example 1

[0084] (1) Planting: Select fertile, moist and well-drained land as planting fields and level the land; after leveling the land, plant 110 plants per mu, apply 10-20 kg of base fertilizer and soil mixture to each planting hole, and transplant the papaya seedlings into the planting holes.

[0085] Furthermore, 30 different red-flowered wrinkled papaya trees of the same age and size were selected from the plantation for an experiment;

[0086] (2) Flower thinning: Flower thinning begins at the initial flowering stage of red-flowered wrinkled papaya.

[0087] The planting hole spacing in step (1) is (1.8~2.2m)×(1.4~1.6m), and the depth is 60~70cm.

[0088] The base fertilizer mentioned in step (1) is well-rotted manure, a mixture of 2000-3000 kg of livestock manure, bedding straw, and feed residue per mu, plus 20-30 kg of superphosphate and 5-10 kg of potassium sulfate (or 30-50 kg of wood ash).

[0089] The thinning of flowers mentioned in step (2) means retaining 3 to 5 flowers in the middle of each inflorescence.

[0090] Pest and disease control, fertilizer and water management, etc. are carried out in accordance with the usual procedures. Harvest when the skin of the red-flowered wrinkled papaya turns greenish-yellow.

[0091] Comparative Example 2

[0092] (1) Planting: Select fertile, moist and well-drained land as planting fields and level the land; after leveling the land, plant 110 plants per mu, apply 10-20 kg of base fertilizer and soil mixture to each planting hole, and transplant the papaya seedlings into the planting holes.

[0093] Furthermore, 30 different red-flowered wrinkled papaya trees of the same age and size were selected from the plantation for an experiment;

[0094] (2) Flower thinning: Flower thinning should begin at the initial flowering stage of the red-flowered wrinkled quince;

[0095] (3) Spraying: Wash 500g of thinned flowers with 20kg of water to collect pollen (about 20g), remove petals and other debris, and spray the flowers once at the beginning of flowering and once at the peak of flowering, with each spray volume being 0.5L;

[0096] The planting hole spacing in step (1) is (1.8~2.2m)×(1.4~1.6m), and the depth is 60~70cm.

[0097] The base fertilizer mentioned in step (1) is well-rotted manure, a mixture of 2000-3000 kg of livestock manure, bedding straw and feed residue per mu, plus 20-30 kg of superphosphate and 5-10 kg of potassium sulfate (or 30-50 kg of wood ash).

[0098] The thinning of flowers mentioned in step (2) means retaining 3 to 5 flowers in the middle of each inflorescence.

[0099] Pest and disease control, fertilizer and water management, etc. are carried out in accordance with the usual procedures. Harvest when the skin of the red-flowered wrinkled papaya turns greenish-yellow.

[0100] Comparative Example 3

[0101] (1) Planting: Select fertile, moist and well-drained land as planting fields and level the land; after leveling the land, plant 110 plants per mu, apply 10-20 kg of base fertilizer and soil mixture to each planting hole, and transplant the papaya seedlings into the planting holes.

[0102] Furthermore, 30 different red-flowered wrinkled papaya trees of the same age and size were selected from the plantation for an experiment;

[0103] (2) Powdering: At the beginning of the flowering period of the red-flowered wrinkled papaya, the flowers are thinned out. The thinned flowers are used to gently rub the open flowers directly to make the pollen stick onto the pistil of the open flowers.

[0104] The planting hole spacing in step (1) is (1.8~2.2m)×(1.4~1.6m), and the depth is 60~70cm.

[0105] The base fertilizer mentioned in step (1) is well-rotted manure, a mixture of 2000-3000 kg of livestock manure, bedding straw and feed residue per mu, plus 20-30 kg of superphosphate and 5-10 kg of potassium sulfate (or 30-50 kg of wood ash).

[0106] The thinning of flowers mentioned in step (2) means retaining 3 to 5 flowers in the middle of each inflorescence.

[0107] Pest and disease control, fertilizer and water management, etc. are carried out in accordance with the usual procedures. Harvest when the skin of the red-flowered wrinkled papaya turns greenish-yellow.

[0108] The yields of the above embodiments and comparative examples are shown in Table 2.

[0109] Table 2. Yield of Red Flower Papaya

[0110]

[0111] Example 9

[0112] The contents of protocatechuic acid, chlorogenic acid, oleanolic acid and ursolic acid in the red-flowered wrinkled papaya obtained in Examples 3-8 and Comparative Examples 1-3 were determined.

[0113] The determination method is as follows:

[0114] 1. Determination of protocatechuic acid and chlorogenic acid content

[0115] 1.1 Chromatographic conditions for constructing the standard curve: The chromatographic column was a Shim-pack CLC-ODS (4.6 mm × 250 mm, 5 μm); the mobile phase was acetonitrile-0.1% phosphoric acid solution (15:85); the detection wavelength was 259 nm for 0-14 min and 325 nm for 14-25 min; the column temperature was 25 ℃; the flow rate was 0.5 mL / min; and the injection volume was 20 μl for analysis.

[0116] Preparation of reference solution: Accurately weigh 10 mg of protocatechuic acid and chlorogenic acid reference standards and place them in a 50 mL volumetric flask. Add a certain amount of 50% methanol aqueous solution, dilute to the mark, and shake well to obtain a reference stock solution with a concentration of 0.2 mg / mL.

[0117] Preparation of standard curves: Accurately measure the stock solution of the reference standard, add an appropriate amount of 50% methanol aqueous solution, and dilute to the mark. Prepare concentrations of 5, 10, 20, 40, 80, 160, and 200 μg / mL. Inject 10 μL of each concentration solution into the HPLC. Measure the peak area of ​​the reference standard under the above chromatographic conditions, and perform regression analysis on the sample concentration (C) and peak area (A). The final regression equations for protocatechuic acid and chlorogenic acid are: Y = 46719X + 17503 (R² = 0.9998) and chlorogenic acid, respectively: Y = 30694X + 19725 (R² = 0.9999). The linear relationship is good in the concentration range of 5-200 μg / mL.

[0118] 1.2 Sample Determination

[0119] Accurately weigh 0.5 g of papaya powder (passed through an 80-mesh sieve) and place it in a stoppered Erlenmeyer flask. Add 25 mL of 50% methanol solution, weigh, and extract ultrasonically for 30 min. Cool to 25℃. Make up the weight loss with 50% methanol, shake well, filter, and collect the filtrate to obtain the test solution. Accurately pipette 10 μl of the sample solution and inject it into the liquid chromatograph for determination. The results are shown in Table 3.

[0120] 2. Determination of Oleanolic Acid and Ursolic Acid Content

[0121] The content of papaya was determined according to the method for determination of papaya content in Part I of the Pharmacopoeia of the People's Republic of China (2020 Edition) [using high performance liquid chromatography (General Rule 0512)]. The Pharmacopoeia stipulates that the total amount of oleanolic acid and ursolic acid in papaya, calculated on a dried basis, shall not be less than 0.5%.

[0122] Chromatographic conditions: Column: Hypersil GOLD Dim (4.6 mm × 250 mm, 5 μm). Mobile phase: methanol-water-glacial acetic acid-triethylamine (265:35:0.1:0.05), flow rate: 1.0 mL / min, column temperature: 18 °C; detection wavelength: 210 nm. Under these chromatographic conditions, the resolution between oleanolic acid and ursolic acid in the sample is greater than 1.5. The theoretical plate number, calculated based on the oleanolic acid peak, should be no less than 5000.

[0123] Preparation of reference solutions: Accurately weigh 10 mg of oleanolic acid and ursolic acid reference standards into 50 ml volumetric flasks, add methanol to prepare reference solutions, which are used as stock solutions.

[0124] Preparation of the test solution: Take about 0.5g of the fine powder of this product, accurately weigh it, place it in a stoppered conical flask, accurately add 25ml of methanol, weigh it, sonicate it (power 250W, frequency 40KHZ) for 20min, cool it, weigh it again, make up the weight loss with methanol, shake well, filter it, and take the filtrate to obtain the test solution.

[0125] Assay method: Accurately pipette 20 μl each of the reference solution and the test solution into the liquid chromatograph and determine the results. The results are shown in Table 3.

[0126] Table 3. Determination of Papaya Content (%)

[0127]

Claims

1. A method for increasing the yield of safflower wrinkled papaya, characterized in that... Includes the following steps: (1) Planting: After leveling the land, dig several planting holes, apply 10-20 kg of base fertilizer and soil mixture to each planting hole, and transplant the papaya seedlings into the planting holes; (2) Applying powder: Start thinning the flowers at the beginning of the flowering period of the red-flowered wrinkled papaya, and use the thinned flowers to gently rub the blooming flowers directly; (3) Spraying pollen: Collect pollen from the thinned flowers, add water extract of Malus hupehensis leaves, and spray it on the flowers once during the initial flowering period and once during the full flowering period. (4) Fruit preservation: Spray the water extract of Hubei crabapple leaves onto the fruit during the initial fruiting stage; The preparation method of the water extract of Malus hupehensis leaves mentioned in steps (3) and (4) is as follows: using Malus hupehensis leaves as raw material, extracting them by heating in a water bath, concentrating them to a phlorizin content of 2.0 wt%, and adjusting the phlorizin content to 0.1-1.0 wt% when using them.

2. The method for increasing the yield of safflower wrinkled papaya according to claim 1, characterized in that: The spacing between rows and plants in the planting holes mentioned in step (1) is (1.8~2.2m)×(1.4~1.6m), and the depth is 60~70cm.

3. The method for increasing the yield of safflower wrinkled papaya according to claim 1, characterized in that: The base fertilizer mentioned in step (1) is one of the farmyard manures mainly composed of well-rotted pen, fence, compost, and biogas residue. It consists of a mixture of 2000-3000 kg of livestock manure, straw and feed residue per mu, plus 20-30 kg of superphosphate and 5-10 kg of potassium sulfate.

4. The method for increasing the yield of safflower wrinkled papaya according to claim 1, characterized in that: The thinning of flowers mentioned in step (2) means retaining 3 to 5 flowers in the middle of each inflorescence.

5. The method for increasing the yield of safflower wrinkled papaya according to claim 1, characterized in that... The base fertilizer mentioned in step (1) is one of the farmyard manures mainly composed of well-rotted pen, fence, compost, and biogas residue. It consists of a mixture of 2000-3000 kg of livestock manure, bedding straw and feed residue per mu, plus 20-30 kg of superphosphate and 30-50 kg of wood ash.

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