CN116918661A - Breeding method of new strain of paeonia lactiflora - Google Patents
Breeding method of new strain of paeonia lactiflora Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/25—Root crops, e.g. potatoes, yams, beet or wasabi
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- Life Sciences & Earth Sciences (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
A breeding method of a new strain of paeonia lactiflora comprises the following steps: s1, establishing a peony germplasm resource database; s2, strain comparison test; s3, strain comparison test; s4 multipoint test: carrying out multi-point tests for a plurality of periods on the new paeonia lactiflora strain, the local hybrid group good strain and the foreign good strain, observing and comparing the weather characteristics, biological properties, yield, quality and resistance of each tested strain/variety, and verifying the yield, specificity, stability and consistency of the new paeonia lactiflora strain; s5 production test: carrying out a production test on the new peony strain, the local hybrid group good strain and the foreign good strain; s6, field technology identification: performing primordial identification on the new paeonia lactiflora; determining AG and MADS2 genes related to stamen of peony petal stamen; s7, evaluating the characteristic characteristics of the new line. The invention adopts a systematic breeding method to obtain a new stable-yield paeonia lactiflora strain with high quality, high yield and strong adaptability.
Description
Technical Field
The invention relates to the technical field of Chinese medicinal material breeding, in particular to a breeding method of a novel paeonia lactiflora line with remarkable appearance character specificity, stable and consistent group character, excellent quality and high yield.
Background
Paull Paeonia lactiflora Pall is a basic plant of radix Paeoniae alba (Paeoniae Radix Alba) and radix Paeoniae Rubra (Paeoniae Radix Rubra) which are commonly used as Chinese medicines. The root of Paeonia lactiflora collected in summer and autumn is cleaned, head, tail and fine root are removed, and the Paeonia lactiflora is boiled in boiling water, and then the skin is removed or peeled, and then the Paeonia lactiflora is boiled and dried in the sun to obtain the Paeonia lactiflora; has the effects of nourishing blood, regulating menstruation, astringing yin, arresting sweating, softening liver, relieving pain, suppressing liver yang, etc., and can be widely used for treating diseases such as blood deficiency, sallow complexion, menoxenia, spontaneous perspiration, night sweat, hypochondriac pain, abdominal pain, limb contracture pain, headache, dizziness, etc. The root, the fibrous root and the sediment of the paeonia lactiflora dug in spring and autumn are removed, and the paeonia lactiflora dug in spring and autumn becomes red paeony root after sun drying; has effects of clearing heat, cooling blood, removing blood stasis and relieving pain, and can be used for treating diseases such as heat-entering nutrient blood, toxic heat, macula, hematemesis, epistaxis, conjunctival congestion, swelling and pain, liver Yu Xie pain, amenorrhea and dysmenorrhea. In actual production, cultivated radix paeoniae alba is mainly processed into radix paeoniae alba, and is a resource plant of radix paeoniae alba.
Modern researches have shown that paeonia lactiflora root, paeonia lactiflora and paeonia lactiflora contain monoterpenes and glycosides, tannins and flavonoids. The paeoniflorin, oxidized paeoniflorin, gallic acid, catechin, 1,2,4,5, 6-pentagalloylglucose, benzoylpaeoniflorin and other components have various pharmacological effects of resisting inflammation, relieving pain, replenishing blood, resisting thrombus, protecting liver, resisting depression, resisting tumor, reducing blood glucose and the like.
The traditional road-region major producing areas of white peony root mainly comprise Sichuan (Zhongjiang and canal county), zhejiang (Strong and safe), and Anhui (Bozhou), and white peony root products are respectively known as Sichuan white peony root, zhejiang white peony root and Bo white peony root. The Sichuan province canal county is taken as a traditional road-region main production area of high-quality white paeony root, has long history, and the produced white paeony root has good quality, has the outstanding characteristics of thick root, fat, hard mass, heavy weight, thick foot, proper yellow and white color, white root section and the like, and is known as silver core white paeony root. White peony root in the county is a genuine medicinal material which is conventionally used in China and used for creating foreign exchange in export, and is once the main raw material of a well-known product of 'Tianfu cola'.
Paeonia lactiflora in the county is planted by planting Paeonia lactiflora with bud heads. Due to a long-term single asexual propagation mode and the variety breeding of medicinal materials in the canal county roads which are not carried out so far, the cultivation population is mixed and degenerated, the production area is seriously atrophic, the wild peony germplasm resources in the production area are not effectively protected, and the stability of the canal county peony yield and quality is seriously affected. Therefore, the problems of fine variety formation of the peony seeds in the county and protection of the wild germplasm resources are needed to be solved.
In the prior art, a method for systematically breeding new peony strains is lacking, and sustainable development of the full industrial chain of Paeonia lactiflora/Paeonia lactiflora in the county is hindered.
Disclosure of Invention
In order to solve the technical problems, the invention provides a breeding method of a new peony strain, which adopts a systematic breeding method to obtain a new stable-yield peony strain with high quality, high yield and strong adaptability.
The invention is realized by the following technical scheme: a breeding method of a new strain of paeonia lactiflora comprises the following steps:
s1, establishing a peony germplasm resource database: establishing a peony germplasm resource nursery and a peony excellent germplasm resource expansion breeding nursery, collecting peony nursery germplasm data, and establishing a peony germplasm resource database;
s2 strain comparison test: screening a plurality of excellent variant single plants from a database, carrying out a plurality of period strain comparison tests on the excellent variant single plants and a local excellent hybrid population, observing the weather characteristics, biological properties, yield, quality and resistance of the single plants, screening a plurality of excellent strains, and purifying the excellent strains to form stable excellent strains;
s3 strain comparison test: carrying out a plurality of cycles of strain comparison tests on stable good strains and local good hybrid populations, observing the climatic period characteristics, biological characters, yield, quality and resistance of the single strain, and screening out new peony strains;
S4 multipoint test: carrying out multi-point tests for a plurality of periods on the new paeonia lactiflora strain, the local hybrid group good strain and the foreign good strain, observing and comparing the weather characteristics, biological properties, yield, quality and resistance of each tested strain/variety, and verifying the yield, specificity, stability and consistency of the new paeonia lactiflora strain;
s5 production test: performing production tests on the new paeonia lactiflora strain, the local hybrid group good strain and the foreign good strain, observing and comparing the characteristics of the weathers, the biological properties, the yield, the quality and the resistance of each tested strain/variety, and verifying the yield, the specificity, the stability and the consistency of the new paeonia lactiflora strain;
s6, field technology identification: performing primitive identification on the new strain of paeonia lactiflora, and identifying the characteristics of plant morphology, flower characteristics, root morphology, color, section characteristics, smell and the like of the new strain; determining AG and MADS2 genes related to stamen petal in paeonia lactiflora new strain and foreign excellent strain by qRT-PCR method;
s7, evaluating the characteristic characteristics of the new strain: and comprehensively evaluating the plant base and morphological characteristics, the fertility characteristics and the biological characteristics of the new variety and the yield and quality of the paeonia lactiflora root bars, wherein the comprehensive evaluation content comprises the plant base and morphological characteristics, the fertility characteristics and the biological characteristics and the yield and quality of the paeonia lactiflora root bars.
Further, in the step of S4 multipoint test and the step of S5 production test, when the biological properties and the biological yield are measured, the biological properties and the biological yield of the overground part and the underground part of the plant are measured respectively.
Further, when the biological characteristics and the biological yield are measured, the yield of the dry product district of the four-year-old paeonia lactiflora root strip of the test strain/variety is converted into the mu yield, and then the statistical, variance and multiple analysis comparison is carried out, wherein the single analysis and the multiple period combination analysis of each period are included.
In the step of S4 multipoint test and the step of S5 production test, when the quality of the four-year-old paeonia lactiflora root of the reference strain/variety is observed and compared, the measured indexes are extract (%) and paeoniflorin (%), and then statistics, variance and multiple analysis are performed, wherein the analysis comprises single analysis and multi-period combined analysis of each period.
Further, stability analysis and fitness analysis are included.
Further, in the step of S4 multipoint test and the step of S5 production test, sandy loam with uniform fertility, deep soil layer, flat land and good drainage is selected in the test land.
Further, in the step of S4 multipoint test and the step of S5 production test, protection rows are arranged on two sides of a field cell, and excellent strains of local hybrid groups are planted.
Further, in the step of S4 multipoint test and the step of S5 production test, the observed data of the morphological feature value of the plant when the biological trait is observed includes: plant height, branch number, flower color, petal, carpel number, plush and leaf characteristics, and root strip appearance characteristic observation data comprise: skin color, section color, presence or absence of wood core and root bar features.
Further, in the step of S2 strain comparison test, the row spacing of the planted strains is 30cm multiplied by 40cm, the planted strains are cultivated in autumn, and the planted strains are picked and excavated in autumn after 3 years, and are randomly arranged; in the S3 strain comparison test step, the row spacing of the planted strains is 30cm multiplied by 40cm, the cell specification is 1.6mmultiplied by 1.5m=2.4 square meters, 5 strains/row multiplied by 4 rows=20 strains are planted in each cell, and the plants are cultivated in autumn, harvested in autumn in 4 years and randomly arranged; in the S4 multipoint test step, the field cells are designed to be 3 multiplied by 3, the field cells are arranged in random groups, the row spacing of the planted plants is 30cm multiplied by 40cm, the specification of the cells is 1.6mX3.0m=4.8 square meters, and 10 plants/row multiplied by 4 rows=40 plants of the planted peonies are arranged in the cells; in the S5 production test, the field cell is designed to be 3 multiplied by 3, the random group arrangement is adopted, the row spacing of the planted plants is 30cm multiplied by 40cm, the cell specification is 4.0mX6.0m=24 square meters, and 20 plants/row multiplied by 10 rows=200 plants of the cell planted Chinese herbaceous peony.
Further, in the steps of the S2 strain comparison test, the S3 strain comparison test, the S4 multipoint test and the S5 production test, the cultivation considerations of the paeonia lactiflora include: the method comprises the steps of producing environment conditions, a cultivation method, a disease and insect comprehensive control method, a harvesting and post-harvesting treatment method, packaging and storage.
Compared with the prior art, the application has the following advantages and beneficial effects: a breeding method of new strain of radix Paeoniae is characterized by establishing a radix Paeoniae germplasm resource database, a strain comparison test, a multipoint test, a production test, a field technology identification, a new strain characteristic evaluation system and other breeding steps, defining specific operation rules of the breeding steps, realizing the standardization and systemization of breeding of new strain of radix Paeoniae, obtaining new strain of radix Paeoniae with high quality, high yield and strong adaptability and promoting sustainable development of regional radix Paeoniae industry.
Drawings
The accompanying drawings, which are included to provide a further understanding of the embodiments and examples of the application and are incorporated in and constitute a part of this specification, illustrate embodiments and examples of the application and together with the description serve to explain the principles of the application. In the drawings:
FIG. 1 is a technical scheme of a breeding method of a new strain of Paeonia lactiflora;
FIG. 2 is a schematic diagram of a field layout of a QBS-1 multi-point test according to the present application;
FIG. 3 is a graph of a first production cycle (2014-2018) multiple test of the inventive channel white No. 1;
FIG. 4 is a graph of a second production cycle (2018-2022) multi-point test of the present application under Trench white 1;
FIG. 5 is a plant view of the present application for a channel white number 1 (QBS-1) and two control varieties (ZBSCK, QSCK) multipoint test;
FIG. 6 is a graph comparing morphological features of three reference lines/variety flowers of the present invention;
FIG. 7 is a graph comparing morphological features of the front of leaves of three reference lines/varieties according to the present invention;
FIG. 8 is a graph comparing morphological features of the back of leaves of three reference lines/varieties according to the present invention;
FIG. 9 is a graph showing the comparison of the morphology of the field plants of the invention, queen No. 1 (left) and Zhong Jiang Baihua (right);
FIG. 10 is a morphological feature of a line plant of line Violet 1 (QBS-1) of the invention;
FIG. 11 is a morphological feature of a Jiang Baihua peony (ZBSCK) control variety plant of the present invention;
FIG. 12 is a graph of analysis of variance of the first cycle (2014-2018) multi-point test yields of the present invention;
FIG. 13 is a graph of analysis of variance of the yield of a second cycle (2018-2022) multi-point test of the present invention;
FIG. 14 is an analysis of variance of the production of a two-cycle (2014-2022) multi-point trial of the present invention;
FIG. 15 is a graph showing the stability (mean-coefficient of variation) analysis of a multi-point test variety according to the present invention;
FIG. 16 is a graph of a multi-point test variety fitness analysis of the present invention;
FIG. 17 is a graph of analysis of variance (extract and paeoniflorin) of the quality of a first cycle (2014-2018) multipoint test of the present invention;
FIG. 18 is a graph of analysis of variance (extract and paeoniflorin) of the quality of a second cycle of production (2018-2022) multi-point test of the present invention;
FIG. 19 is a graph of analysis of variance (extract and paeoniflorin) of the quality of a multi-point trial of two consecutive production cycles (2014-2022);
FIG. 20 is a graph of stability analysis of quality of a two-cycle (2014-2022) multi-point test in succession in accordance with the present invention;
FIG. 21 is a graph showing the stability (fitness) analysis of the quality of a two-cycle (2014-2022) multi-point test in series according to the present invention;
FIG. 22 is a schematic diagram of a field layout of QBS-1 production test of Paeonia lactiflora in county of the invention;
FIG. 23 is a graph of analysis of variance of the yield of the production test of the present invention (2018-2022);
FIG. 24 is a graph of stability analysis of the yield of the cultivar of the production test of the invention (2018-2022);
FIG. 25 is a graph showing the stability (fitness) analysis of the yield of the strain varieties of the production test (2018-2022) of the present invention;
FIG. 26 is a graph of analysis of variance of the quality of the production test of the present invention (2018-2022);
FIG. 27 is a graph of stability (mean-CV) analysis of the quality of the production test of the present invention (2018-2022);
FIG. 28 is a graph of stability (fitness) analysis of the quality of the production test of the present invention (2018-2022).
Detailed Description
The invention is further described below with reference to examples. The following embodiments are merely specific examples of the present invention, but the design concept of the present invention is not limited thereto, and any insubstantial modification of the present invention by using the concept shall belong to the behavior of infringement of the protection scope of the present invention.
In this embodiment, the selection and breeding process of the new strain QBS-1 is described in detail, and the breeding technology route diagram is shown in FIG. 1.
1. Establishing a peony germplasm resource database
By means of focus, investigation of producing area and other forms, 107 parts of wild and cultivated paeonia lactiflora germplasm resource materials of Sichuan (Zhongjiang and canal county), zhejiang, anhui, shanxi, shandong and the like are collected. Wherein, 20 parts of Chinese herbaceous peony, 48 parts of canal-county paeonia lactiflora, 12 parts of Zhejiang paeonia lactiflora, 15 parts of Anhui paeonia lactiflora, 6 parts of Shanxi paeonia lactiflora and 6 parts of Shandong paeonia lactiflora. And establishing a peony germplasm resource nursery and a peony excellent germplasm resource expanding breeding nursery in the canal county. Meanwhile, a wild germplasm resource protection area of paeonia lactiflora is also established in the canal county. From the mixed population of the traditional Chinese herbaceous peony cultivated in the Shangzhen village of the Tongzhenhan, the breeding unit personnel systematically select 12 variant single plants with excellent germplasm resources through observation and comparison of the single plants for many years.
2. Strain comparison test
On the basis of initially selecting and collecting and sorting 12 excellent variation breeding materials of single plants of the Paeonia suffruticosa in the county, a strain comparison test (row spacing of planted plants is 30cm multiplied by 40cm, planting in autumn, digging in autumn after 3 years, random group) of two continuous production cycles is carried out by taking QSCK (white flowers are taken as main, pink flowers are taken as main) of a traditional cultivation mixed population in a main production area of the Paeonia suffruticosa in the county to observe the characteristics of the single plants in the weather, biological characters, yield, quality, resistance and the like of the single plants, and 3 excellent strains (QBS-1, QBS-2 and QBS-3) of the Paeonia suffruticosa in the county are determined. Meanwhile, the excellent strain materials are widely collected in the area of the Paeonia lactiflora and property in the county, the resource group is expanded, the breeding is carried out, and the stable strain is formed by purification.
3. Comparative test of strain
In 2006-2014, 3 peony-producing villages such as Qinggui dam, tianshan of the middle beach and the like in the Qinggai-Baba garden in the county with the mixed community of Paeonia lactiflora (QSCK) as a control variety were subjected to strain comparison tests of the 3 excellent strains (QBS-1, QBS-2 and QBS-3) in two continuous production cycles (row spacing of the planted strains is 30cm multiplied by 40cm, cell specification is 1.6mmultiplied by 1.5m=2.4 square meters, 5 Paeonia lactiflora/row multiplied by 4 row=20 plants are planted in each cell, autumn cultivation is performed, and excavation is performed in autumn of 4 years, and random granules are formed). Meanwhile, the weather-proof property, biological property, yield, quality and resistance of each test strain are examined in the field, and a line of QBS-1 is determined.
Meanwhile, the excellent single plant germplasm resource materials with more than one excellent are widely collected in the main production area of the Paeonia lactiflora in the canal county, expanded breeding is carried out, a multipoint comparison test in the next stage is prepared, and the Paeonia lactiflora (ZBSCK) of Jiang Baihua which is traditionally cultivated in the main production area of the Jiang Queyao canal in the planting is introduced.
4. Multipoint test for two consecutive production cycles
In 2014-2022, two production cycle multi-point tests were performed on 3 reference lines/varieties such as canal white 1 (QBS-1) line, control variety canal county peony hybrid population (QSCK) and control variety Jiang Baihua peony (ZBSCK) in canal county and other county, qing county Gui-Cheng Yuan village, zhongjun Tianshan village, he Xiao mountain village, dong' an Xiang Jiu village and the like in the main production area of the peony road. The field cells are designed to be 3 multiplied by 3, the random block arrangement is realized, the row spacing of the planted plants is 30cm multiplied by 40cm, the cell specification is 1.6mX3.0m=4.8 square meters, and 10 plants/row multiplied by 4 rows=40 plants of the planted Chinese herbaceous peony are planted in the cells.
Multi-point test of the first production period is carried out in 2014-2018, autumn cultivation is carried out, and harvesting is carried out in autumn of 4 years;
the multi-point test of the second production period is carried out in 2018-2022, the cultivation is carried out in autumn, and the harvesting is carried out in autumn of 4 years.
The second production cycle multipoint test non-harvested plants were used for field technical identification in the spring 2023.
The yield, specificity, stability and consistency of the line QBS-1 were verified by observing and comparing the climatic characteristics, biological traits, yield, quality and resistance of each of the test lines/varieties.
4.1 test overview
1) Purpose of test
Multiple point comparison tests of regional ecological adaptability of the canal white No. 1 (QBS-1) strain in two continuous production periods are carried out in main areas of Paeonia in Sichuan county and in 2018-2022 to evaluate the performances of adaptability, stable yield, high yield, stress resistance, root strip quality and other important characteristics of the canal white No. 1 strain in the ecological area of the Daba mountain.
2) Test period
2014-2018, a first production cycle 4 years multi-point trial;
years 2018-2022, the second production cycle 4 years of multipoint test.
3) Test site
And (3) arranging a multipoint test in a suitable ecological area of the Daba mountain centering on the main production villages and towns of the main production area of the traditional road region of the Paeonia lactiflora in the canal county. Multiple trials of line white number 1 (QBS-1) for two consecutive production cycles, at years 2014-2018 and 2018-2022, were each selected at the same 5 trial points, as shown in table 1.
Table 1 geographical information table for each test point of strain number 1 in the canal
Note that: the multipoint experiments of two continuous production periods are all selected at the same place, and the place is the scientific and technological key country of the main product of paeonia lactiflora and province.
4) Reference strain/variety
(1) QBS-1 strain is a canal white No. 1 strain bred from Paeonia lactiflora in canal county. And before 2014, the breeding unit breeds a high-quality high-yield white-flower imperial crown-shaped heavy-petal excellent single plant from a mixed population of Chinese herbaceous peony planted in a main production area of a canal county and a road and collecting local wild germplasm resource materials in a systematic way according to the morphological characteristics and the fertility characteristic differences of plants, passes a plant line comparison test, and is further purified and cultivated into an excellent line with stable shape.
(2) The QSCK control variety is a mixed population of traditional planting in a main production area of a canal county paeonia lactiflora, and mainly comprises two types of mixed white peony, pink peony and the like. Wherein white peony is the main germplasm resource type of the mixed population of Paeonia lactiflora in the county.
(3) The ZBSCK contrast variety is Jiang Baihua Paeonia lactiflora, and is one of the traditional main cultivars in the main production area of Jiang Dao. The variety passes field technical identification of provincial experts in 2019, and provincial grade 2020 is identified as a new white flower Chuan white peony root CBS-B variety.
5) Design of experiment
And 5 peony main producing villages (town) of 2014-2018 and 2018-2022 are respectively subjected to multi-point tests of two continuous production periods in canal county Linba-ma garden, qing Gui dam, tianshan mountain in the middle beach, xiao mountain, dong' an Guangqiao and the like.
And selecting sandy loam with characteristics of representativeness, uniform fertility, deep soil layer, flat land and good drainage, and adopting a land block with consistent front working as a test land. The area test design was 3×3, using a one-way random block arrangement, as shown in fig. 2. The test line/variety 3 (1 line, 2 control variety), the test was repeated 3 times (n=3). The planting specification of the paeonia lactiflora is designed to be 30cm x 40cm, the planting density is 5500 plants/mu, the test cell is a rectangular high box, 1.6mx3.0m=4.8 square meters, and 10 plants/row x 4 rows=40 plants of the paeonia lactiflora are planted in each cell, and the cells are planted in staggered mode. Setting a protection row, and planting a canal county peony mixed population (QSCK).
The first production cycle of multipoint test was planted in autumn 2014 (10 months), and the peony plants were harvested after withering in autumn 2018 (10 months). The multipoint test of the second production period is carried out in 2018 autumn (10 months) after the peony plants in 2022 autumn wilt (10 months) are planted in the same test point. And 20 plants are collected and detected in each cell. The multipoint test points of the line 1 and the control line are shown in fig. 3 and 4.
6) Field management and observation and measurement
(1) Management gist
According to the technical regulations for producing white peony root (DB 51/T805-2008), the preparation of soil, fertilization, planting, intertillage weeding, pest control, harvesting and processing are carried out.
(2) Measurement by observation
And (5) observing the plant waiting period of each test strain/variety, and recording the growth vigor and uniformity of the plants. The annual growth period starts from the rapid germination and growth (2 months) of buds of the paeonia lactiflora plants in the dormant period, and the seedlings are withered and fallen (10 months) after stem and leaf extraction, flower bud, flowering and fruiting. The annual dormancy period starts from the withered seedling of the paeonia lactiflora plant (10 months) to the germination and growth of the bud of the paeonia lactiflora plant (2 months) in the next year.
(3) Biological traits and biological yield investigation
The four-year-old plants of the test strain/variety enter the full-bloom period (4-5 months), and the biological characters and the biological yield (g/plant) of the plants are regulated; in autumn (10 months > harvest period), cell yield measurements (kg/cell) were performed.
In each cell of each test line/variety, 5 representative plants were randomly selected, and whole plants were harvested, i.e., the number of representative plants harvested per test line/variety was 5 plants/cell×3 cells=15 plants (n=15). And (5) preserving the picked whole plants, transporting the whole plants back to the room, and respectively measuring the biological properties and the biological yield (g/plant) of the overground parts and the underground parts of the plants. The biological properties of the aerial parts are measured by plant height, crown width, branch number, leaf number and morphology, flower number, flower color and corolla morphology, etc. The biological characteristics of the underground part are mainly measured by root depth, root width, root number, circumference of the reed, bud number, morphological characteristics and the like. The main measurement indexes of biological yield include (1) plant total biomass and its yield constitution (aerial part has leaf weight, stem weight, flower weight, etc., underground part has root weight, top grade root weight, bud weight, etc.). (2) Fresh product yield and dry product yield, and calculate the dry rate (%), etc.
(4) Root appearance observation
Taking the root strips of the four-year-old paeonia lactiflora plants of the acquired test strain/variety as an observation material, and observing the surface color, the section color, the lignification, the pest hazard degree and the like of the root strips after naturally drying in the shade. According to the size of the paeonia lactiflora root strips, the paeonia lactiflora root strips are classified into an upper grade type and an equal grade type.
(5) Determination of root strip yield
Four-year-old plants of the multi-point test reference strain/variety are collected and tested in autumn by taking the district of each test point as a measurement unit. 20 plants were randomly picked from each plot, soil and aerial parts were removed, root fresh weights were measured, and the average fresh weights of the individual plants (20 total fresh weights of root bars kg/20 plants kg/plant) and the average fresh weights of the plot (20 total fresh weights of root bars kg×40 plants/20 plants kg/plot) were calculated.
And (3) respectively placing the collected and tested fresh root strips of the paeonia lactiflora in a cool and ventilated place for natural drying in the shade, and measuring and recording the dry product weight (kg/cell) of the root strips of the paeonia lactiflora in each test cell. At the same time, the rate of drying of paeonia lactiflora root strips (20 total dry weight kg/total fresh weight kg x 100% of the cell) was calculated.
The dry product yield of the four-year-old paeonia lactiflora root stripes of the reference strain/variety is respectively converted into the district yield (kg/district, 40 strains) and the acre yield (kg/acre). The yield per mu of 5500 plants planted in each mu is kg multiplied by 5500/40 of the yield of the district. And carrying out statistics and variance analysis on acre yield of four-year-old paeonia lactiflora root dry weight of each test strain/variety in the multipoint test.
(6) Mass analysis
According to the method and standard under the items of white peony root and red peony root in the Chinese pharmacopoeia (reference in the early 2015 edition and in the later 2020 edition, one part) the dry weight samples of the root bars of each cell of the two production cycle multipoint tests of 3 test lines/varieties are measured, and meanwhile, the dry weight samples of the root bars of each test cell are randomly sampled and sent to the Chinese medicine research institute of southwest traffic university for measurement.
7) Processing and statistical analysis of test data
The dry weight yield (kg/cell), the converted mu yield (kg/mu), the extract (%) of the root and paeoniflorin (%) of the root of each cell of the 3 test lines/variety multipoint test are expressed as an average value (x+ -SD), and the statistical test and analysis are performed on each relevant data.
4.2 test results and analysis
1) Comparison of the weathering characteristics of the various test lines/varieties in the multipoint test
The observation of the characteristics of each of the test lines/varieties in the period of the multi-point test of two consecutive production cycles of 2014-2018 and 2018-2022 was carried out, and the results are shown in Table 2.
TABLE 2 comparison of the characteristics of the candidate lines/varieties in the main production area of Paeonia lactiflora
Note that: the dates in the table are the average of observations of the test line/variety in each test point of a multi-point test for two consecutive production cycles.
Compared with the QSCK control varieties of the mixed populations of Chinese Paeonia lactiflora ZBSCK and Paeonia lactiflora in China Jiang Baihua, the QBS-1 strain has the advantages of long annual growth period, fast emergence and leaf expansion, strong branching power and long flowering period.
2) Morphological feature comparison of each reference strain/variety plant in multipoint test
Morphological characteristics of four-year-old plants of each test-point reference strain/variety are observed in a multi-point test of two continuous production periods. Comparison of three reference lines/variety peony representative plants, flowers, leaves is shown in fig. 5-8. A comparison of representative plants and root systems of the line QBS-1 and the control variety ZBSCK of Paeonia lactiflora in China Jiang Baihua is shown in FIGS. 9-11. Morphological feature values of each of the test lines/variety plants are shown in table 3. The results of the root strip appearance property inspection are shown in table 4.
TABLE 3 statistics of main morphological characteristics of each test line/variety plant in the main production area of Paeonia lactiflora in county
Note that: the data in the table are the average of the main morphological characteristics of each test point of the multi-point test reference strain/variety plant for two consecutive production cycles. b statistics n=5×3×5×2.
TABLE 4 appearance characteristics questionnaire for each of the test lines/variety root strips in the main production area of Paeonia lactiflora
As can be seen from tables 3 and 4, compared with the QSCK control varieties of the mixed population of the Chinese Paeonia ZBSCK and the canal Paeonia county of Jiang Baihua, the main morphological characteristics of the representative plants of the canal white No. 1 strain are represented by the high plant size, the number of branches of stems, the thick and strong root, the heavy white flower petal, the stamen petal crown shape and the like. The character characteristics lay a good photosynthetic ecological and nutritional ecological foundation for the high quality and high yield of the canal white No. 1 strain.
The test results show that the climatic period and the plant biological properties of each test strain/variety are stable (P > 0.05) in the multi-point test of 5 test points and two continuous production periods. The canal white No. 1 strain bred from the canal county paeonia district main production area system has obvious specificity, stability and consistency.
3) Results and analysis of multi-point test yield statistics
(1) Multi-point test yield measurement statistical result
Cell yields (kg/cell) of each of the reference lines/varieties of paeonia lactiflora root trunks are tested at two continuous production cycles in 2014-2018 and 2018-2022, and are shown in tables 5 and 6, and converted into yields per mu (kg/mu), and are shown in tables 7 and 8.
The results show that the yields of each of the test lines/varieties of paeonia lactiflora root bars between 3 replicates at 5 test points of two consecutive production cycles have no significant difference (P > 0.05); the yield of paeonia lactiflora root stripes among the three test lines/varieties has very significant variability (P < 0.001). The root yield of the three test lines/varieties was in the order from high to low: canal white number 1 (QBS-1), medium Jiang Baihua peony (ZBSCK), canal county peony mixed population (QSCK). The yield of Paeonia lactiflora root bars of the line QBS-1 (QBS-1) is extremely higher than those of Paeonia lactiflora (ZBSCK) of medium Jiang Baihua and Paeonia lactiflora mixed population (QSCK) (P < 0.001). Comparing the peony yields of each test point and each repetition of the two production cycle multipoint tests, the peony yield of the first production cycle (2014-2018) is higher than the peony yield of the second production cycle (2018-2022), and the main reason is that the extremely high temperature drought in 2022 leads to the reduction of the overall yield of the peony root strips of the second production cycle (2018-2022) multipoint test.
Table 5 cell yield statistics for the first production cycle (2014-2018) multipoint test
Note that: the data in the table are statistical values of the collection and yield measurement of the four-year-old paeonia lactiflora root strip autumn of each test point of the multi-point test of the first production period. b Increase the yield value for QBS-1 over ZBSCK; c the yield value is increased for QBS-1 over QSCK.
Table 6 the first production cycle (2014-2018) multipoint test translates into a mu yield statistics table
Note that: the data in the table are statistical values of the collection and yield measurement of the four-year-old paeonia lactiflora root strip autumn of each test point of the multi-point test of the first production period.
Table 7 cell yield statistics for the second production cycle (2018-2022) multipoint test
Note that: the data in the table are statistical values of the collection and yield measurement of the four-year-old paeonia lactiflora root strip autumn of each test point of the multi-point test of the second production period. b Increase the yield value for QBS-1 over ZBSCK; c the yield value is increased for QBS-1 over QSCK.
TABLE 8 conversion of the second production cycle (2018-2022) multipoint test into a mu yield statistics
Note that: the data in the table are statistical values of the collection and yield measurement of the four-year-old paeonia lactiflora root strip autumn of each test point of the multi-point test of the second production period.
(2) Analysis of variance and multiple comparisons of first cycle multipoint test yields
Analysis of variance of peony yields from 5 trials 3 lines/cultivars were compared with multiple at 2014-2018 in the first production cycle of multipoint trials (as shown in fig. 12). The results show that the variance analysis of the multi-point test yields in the first production cycle shows that the difference in peony yield (f= 14402.35) between 3 test varieties is significantly greater than the interaction effect (f=0.45) between 5 test sites (f=20.88) and test variety-test sites. The average yield of the root bars of the QBS-1 is 1053.53 kg/mu, which is 7.55 percent and 28.87 percent higher than that of the root bars of Jiang Baihua (ZBSCK, 979.60 kg/mu) and the mixed group of Paeonia in the canal county and the Paeonia in the control variety (QSCK, 817.53 kg/mu), respectively, and the yield is 100 percent.
(3) Analysis of variance and multiple comparisons of second cycle multipoint test yields
Analysis of variance and multiple comparisons of peony yields from 5-site 3-test lines were performed at multiple points during the second production cycle of 2014-2018 (as shown in fig. 13). The results show that the variance analysis of the multi-point test yields in the second production cycle shows that the difference in peony yield (f= 10936.88) between 3 test varieties is significantly greater than the interaction effect (f=5.31) between 5 test sites (f=23.57) and test variety-test sites. The average yield of the root bars of the QBS-1 is 992.80 kg/mu, which is increased by 8.68 percent and 30.14 percent respectively compared with the average yield of the root bars of Jiang Baihua (ZBSCK, 913.53 kg/mu) and the mixed population of Chinese herbaceous peony (QSCK, 762.87 kg/mu) in the control variety, and the yield is increased by 100 percent.
(4) Yield analysis of variance and multiple comparisons of continuous two production cycle multipoint tests
Analysis of variance and multiple comparisons of peony yields from 5-site 3-test lines were performed in two consecutive production cycles at multiple points in 2014-2022 (as shown in FIG. 14). The results show that the variance analysis of the multi-point test yields of two continuous production cycles shows that the difference of the paeonia lactiflora root-bar yields (f= 1725.8) between 3 test varieties is significantly larger than the interaction effect (f=2.77) of 5 test sites (f=35.77) and test variety-test site-year. The average yield of the root bars of the QBS-1 is 1023.17 kg/mu, which is 8.09 percent and 29.48 percent higher than that of the root bars of Jiang Baihua (ZBSCK, 946.57 kg/mu) and the mixed group of Paeonia in the canal county (QSCK, 790.20 kg/mu) in the control variety, respectively, and the yield is 100 percent.
The main reason for the yield reduction of the paeonia lactiflora root stripes in the second production cycle compared with the first production cycle is that the 2022 ultra-large continuous high-temperature drought exists.
Analysis of variance of average yield of root bars of each test strain/variety paeonia lactiflora is shown in Table 9. Multipoint test of two continuous production periods (2014-2022), wherein each test strain/variety of paeonia lactiflora is in the same production period, and different test points have no obvious influence on plant growth and development, biological properties and root yield (P > 0.05). However, the yields of each of the test lines/varieties of paeonia lactiflora root stripes have very significant differences (P < 0.001). The average yield of the root bars of Paeonia lactiflora in two continuous production cycles of a line of GanBai No. 1 (QBS-1) bred from a main production area of a canal county district is 1023.17 kg/mu, which is 8.09% and 29.48% higher than the average yield of the root bars of Jiang Baihua Paeonia lactiflora ZBSCK (946.57 kg/mu) in a control variety and the average yield of the root bars of Paeonia lactiflora QSCK in a canal county Paeonia lactiflora hybrid group (790.20 kg/mu), respectively (P < 0.001). Extreme abnormal climates have a significant impact on each of the test lines/varieties at different test points in the same production cycle.
TABLE 9 average yield of paeonia lactiflora root stripes in two consecutive production cycles (2014-2022) multipoint test
Note that: the second period (2018-2022) in the table leads to reduced yield of root strips of various reference lines/varieties of paeonia lactiflora due to drought at a high temperature of 2022.
(5) Stability analysis of Multi-Point test yield
Stability analysis and fitness analysis (fig. 15 and 16) of the yield of each of the test lines in two consecutive production cycle multipoint tests (2014-2022), the coefficient of variation CV (3.171% -3.842%) of the root bar yield (kg/mu) of paeonia lactiflora in and between each of the test lines/varieties was small, indicating that the root bar yield of each of the test lines/varieties in the multipoint test was relatively stable. Analysis of the fitness of each of the test lines/varieties in the multi-point test shows that the fitness of QBS-1 and ZBSCK reaches 100%.
4) Results of quality detection and statistical analysis of paeonia lactiflora root strips in multi-point test
(1) Detection result of quality of paeonia lactiflora root strips in multipoint test
Two continuous production cycles (2014-2022) are carried out for multipoint tests, and main quality indexes measured by each test strain/variety paeonia lactiflora root strip are extract (%) and paeoniflorin (%). Three test line/variety paeonia lactiflora root strip samples of 2 batches in succession in 2018 and 2022 were measured; and the three test lines/variety paeonia lactiflora root strip samples collected in 2022 are sent to the southwest university of transportation traditional Chinese medicine research institute for determination. The contents of the sample extract and paeoniflorin are shown in Table 10.
Table 10 Multi-Point test peony quality inspection statistics (n=3×2) for two consecutive production periods (2014-2022)
Note that: the data in the table are combined statistical values of quality indexes of four-year-old radix paeoniae alba root detection of each test point of the multi-point test of two continuous production periods. b is a QBS-1 to ZBSCK upgrading value; c is QBS-1 and QSCK.
The result shows that the root bar of the canal white No. 1 (QBS-1) bred by the system of the main production area of the canal county and the district is 24.62 percent and 35.72 percent higher than that of the root bar of Jiang Baihua Chinese herbaceous peony ZBSCK (authentication variety code CBS-B) and the mixed group QSCK of the canal county and the Chinese herbaceous peony respectively through continuous two production periods (2014-2022 years) multipoint experiments; the paeoniflorin content is 33.57% and 9.77% respectively. There is a very significant difference (P < 0.001) between canal white No. 1 and medium Jiang Baihua peony ZBSCK and canal county peony heterogeneous population QSCK.
(2) Analysis of variance and multiple comparisons of quality of root bars of paeonia lactiflora in multipoint test
Two production cycle multipoint tests were continued, and analysis of variance and multiple comparison results of the quality of the paeonia lactiflora root strips of each of the test lines (fig. 17 to 18) were obtained.
The first production cycle (2014-2018) multi-point test shows that the variance analysis and multiple comparison analysis of the index component content of the root bar of the 3 reference lines/varieties at 5 test points show that the extract (%) and paeoniflorin (%) content variance analysis F values (1235.03, 73.50) between the reference lines/varieties have significant differences. The average extract content (35.73%) of the canal white No. 1 extract is 25.23% and 36.39% higher than the content (28.53%) of Jiang Baihua peony ZBSCK and the content (26.20%) of QSCK of the mixed group of canal county peony in the control variety. The content of paeoniflorin No. 1 in the canal white (3.93%) is 31.11% and 13.46% (P < 0.001) higher than the content of Jiang Baihua paeonia ZBSCK in the control variety (3.00%) and the content of QSCK in the mixed population of the canal county paeonia (3.47%), respectively, and the increment point is 100%.
The second production cycle (2018-2022) multi-point test, analysis of variance and multiple comparison analysis of the index component content of root bars of 3 reference lines/varieties at 5 test points showed that the extract (%) and paeoniflorin (%) content analysis of variance F values (2129.02, 175.00) were significantly different between reference lines/varieties. The average content (35.73%) of the extract of GanBai 1 root strip is 23.79% and 35.70% higher than the content (28.87%) of Jiang Baihua peony ZBSCK and the content (26.33%) of QSCK of the mixed group of Paeonia in the control variety. The average content (4.00%) of the paeoniflorin in the canal white No. 1 root is 33.33% and 9.09% (P < 0.001) higher than the content (3.00%) of Jiang Baihua paeonia ZBSCK in the control variety and the content (3.67%) of QSCK in the mixed population of canal county paeonia, respectively, and the increase point reaches 100%.
FIG. 18 analysis of variance of quality (extract and paeoniflorin) for a second production cycle (2018-2022) multipoint test
Two consecutive production cycles (2014-2022) multipoint tests, analysis of variance of the index component content of the root strip of 3 reference lines/varieties at 5 test points, and multiple comparison analysis showed that the extract (%) and paeoniflorin (%) content analysis of variance F values (3434.37, 91.00) between reference lines/varieties were significantly different. The average content of extract of QBS-1 (QBS-1) was 24.51% and 36.04% higher than the content of QSCK (28.70%) of Jiang Baihua Paeonia lactiflora and the content of QSCK (26.27%) of the mixed population of Paeonia lactiflora. The average content (3.97%) of paeoniflorin No. 1 in the canal is 32.22% and 11.21% (P < 0.001) higher than the content (3.00%) of Jiang Baihua paeonia ZBSCK in the canal and county paeonia hybrid population QSCK respectively, and the increment point is 100%.
(3) Stability analysis of quality of paeonia lactiflora root strips in multipoint test
Stability analysis of quality of each of the test lines/varieties of paeonia lactiflora root bars in two continuous production cycle multipoint tests shows that (shown in fig. 20), extract stability of the paeonia lactiflora root bars of different test lines/varieties is represented by the mixed population of (QSCK, cv=1.001%) > Jiang Baihua paeonia lactiflora (ZBSCK, cv=1.277%); stability of paeoniflorin was measured as Jiang Baihua peonies (ZBSCK, cv=0.000%) > canal white No. 1 (QBS-1, cv=2.657%) > canal county mixed population of peonies (QSCK, cv= 13.254%). The quality of the strain No. 1 of the open channel is stable.
(4) Adaptability analysis of quality of paeonia lactiflora root strips in multipoint test
The results of the fitness analysis of the quality of each test line/variety of paeonia lactiflora root strips in two continuous production periods and multipoint tests show that the fitness of the extract of the paeonia lactiflora root strips of the line QBS-1 (QBS-1) in canal white reaches 100% at different test points; the adaptability of the paeoniflorin content of Jiang Baihua paeonia (ZBSCK) and the root strips of the canal county paeonia mixed population (QSCK) at 5 test points is 50%; the adaptability of the extract of the root strips of Paeonia lactiflora (ZBSCK) and paeoniflorin content of Jiang Baihua in the culture medium is 0 in 5 test points. The open channel white No. 1 (QBS-1) strain is a good quality peony germplasm resource suitable for Daba mountain.
4.3 Multi-Point test nodelets
The observation and statistical analysis results of the multipoint experiments of two continuous production periods of 2014-2018 and 2018-2022 show that the growth and development characteristics of 3 reference strains/varieties selected from main production areas of the county and the region Jiang Queyao have significant differences (P < 0.001) in biological characteristics, root strip appearance characteristics, yield and quality of four annual plants. The comprehensive evaluation of each of the test lines/varieties in the multipoint test is as follows:
1) Line QBS-1
The annual growth period of QBS-1 strain is about 260 days, the seedling emergence and leaf expansion are fast, the plant height is 40-100cm, the average branches of single plant is 15.54+/-2.49, the root is thick, the flowering period is long, the white flower is heavy, the stamen is petaloid, the carpel number (3-4) is more than 4, and the villus is present. The average yield of the root strips of the paeonia lactiflora in the multipoint test is 1023.17 kg/mu (8.09% and 29.48% higher than the average yield of Jiang Baihua and canal county paeonia lactiflora mixed groups in a control variety respectively), the average extract content is 35.73% (24.51% and 36.04% higher than the average content of Jiang Baihua and canal county paeonia lactiflora mixed groups QSCK respectively), and the average paeoniflorin content is 3.97% (32.22% and 11.21% higher than the content of Jiang Baihua and canal county paeonia lactiflora mixed groups QSCK respectively). The yield and quality are stable.
2) Control variety of Paeonia lactiflora (ZBSCK) of medium Jiang Baihua
The annual growth period of the medium Jiang Baihua paeonia lactiflora ZBSCK control variety is about 240 days, the plant is 40-70cm high, the average branches of a single plant are 12.73+/-3.61, the root is thick, the flowering period is longer, the flowers are white, the stamen valve chrysanthemum flowers are shaped, and the number of carpel (3-4) is 3 more and fluff exists. The average yield of the paeonia lactiflora root strips in the multipoint test is 946.57 kg/mu, the average content of extract is 28.70%, and the average content of paeoniflorin is 3.00%; the yield and quality are stable.
3) The annual growth period of the QSCK control variety of the QSCK hybrid population of the Paeonia suffruticosa hybrid population (QSCK) control variety of the Paeonia suffruticosa hybrid population of the Paeonia suffruticosa county is about 250 days, the plant height is 40-100cm, the average branches of the individual plant is 10.98+2.21, the roots are thicker, the flowering period is longer, white flowers and pink are mixed, the number of the pericarp is more than 3, and the white flowers have villus and pink does not have villus. The average yield of the paeonia lactiflora root stripes in the multipoint test is 790.20 kg/mu; the average content of extract is 26.27%, and the average content of paeoniflorin is 3.57%; the yield and the quality are more stable. The QBS-1 strain is the white peony in the county, the growing period and the flowering period are longer, the seedling emergence and leaf expansion are quick, the plant is high and the branching force is strong; the root strips are more and thicker, the yield is high, the quality is good (the extract and the paeonia lactiflora back content are high), and the Chinese pharmacopoeia regulation is met.
5. Production test for one production cycle (2018-2022)
In 5 villages of the multipoint test in 2018-2022, a production test of one production cycle was performed by taking the canal white number 1 (QBS-1) strain, the control variety canal county peony hybrid group (QSCK) and Jiang Baihua peony (ZBSCK) in the control variety as reference strains/varieties.
Cultivating in autumn, and harvesting in autumn in the fourth year. The field design and layout scheme are the same as that of the multipoint test, and the difference is that the area of the field and the number of the peonies planted in the field are enlarged, namely, the field is designed to be 3 multiplied by 3, the field is arranged in a random group, the row spacing of the planted plants is 30cm multiplied by 40cm, the cell specification is 4.0mmultiplied by 6.0m=24.0m, and the number of peonies planted in the field is 20 plants/row multiplied by 10 rows=200 plants. Observations and comparisons of the characteristics of the weather, biological traits, yield, quality, resistance and the like were made for each of the test lines/varieties, and the "trisection" of the line QBS-1 was further confirmed.
5.1 purpose of test
On the basis of the first production cycle multipoint test in 2014-2018, the second production cycle multipoint test in 2018-2022 is carried out, and meanwhile, production tests are carried out on 3 test lines/varieties in 5 test points, so that the characteristics of high yield, high quality, stable yield, adaptability and the like of the bred line QBS-1 strain planted in a large area in a suitable ecological area of the Daba mountain can be further evaluated, and scientific basis is provided for field technology identification, application and popularization of the new line QBS-1 strain.
5.2 test protocol
The production test scheme of the QBS-1 strain and the test site, the reference strain/variety, the test design, the field management, the observation and measurement and the test data of the multipoint test are completely the same as those of the statistical analysis scheme, and the difference is that the cell area (24.0 square meters) of the production test is enlarged by 5 times compared with that of the multipoint test (4.8 square meters). 200 plants/cell of the paeonia lactiflora in the production test, and 40 plants/cell of the paeonia lactiflora in the multipoint test. The production test has the following specific scheme:
1) Test time
2018-2022. The peony bud head planting is planted in autumn in 2018, and the peony bud head planting is harvested in autumn in 2022.
2) Test site
Five villages (town) such as Linba hemp garden, qinggui dam, tianshan mountain of the middle beach, xiaoshan, eastern Angustification bridge and the like in main production area of Paeonia area in canal county. Geographical information such as test place name, altitude, longitude and latitude is shown in table 1.
3) Reference strain/variety
Production of 3 tested lines/varieties. The new strain of QBS-1 is named as the new strain of QBS-1, and the control varieties are medium Jiang Baihua Paeonia lactiflora (ZBSCK) and canal county Paeonia lactiflora mixed population (QSCK). Information about each test line/variety is detailed in the multipoint test section of this report.
4) Test design
The area test was designed 3×3, and the test was repeated 3 times (n=3) using a one-factor random block arrangement (see fig. 22); the test cell is a rectangular high box with the specification of 4.0m multiplied by 6.0 m=24.0 square meters; the planting specification is 30cm multiplied by 40cm,4 rows are cultivated by staggered nest, 20 plants/row multiplied by 10 rows = 200 plants are planted in a district, and the planting density is 5500 plants/mu. And planting a canal county peony mixed population (QSCK) in the protection row.
5) Field management and observation and measurement
According to the technical regulations for producing white peony root (DB 51/T805-2008), the preparation of soil, fertilization, planting, intertillage weeding, pest control, harvesting and processing are carried out.
The method and the index of field observation record, biological character and biological yield investigation, root appearance character investigation, root strip yield measurement, quality detection and the like are the same as those of the multipoint test scheme. The weather characteristics, biological properties, yield, quality and resistance of each test strain/variety were examined in the field.
6) Processing and statistical analysis of test data
100 four-year-old paeonia lactiflora root strips are dug from each district, and the yield and the quality are measured. The indexes of the production test paeonia lactiflora root strips (dry products) such as measured yield (kg/cell), converted mu yield (kg/mu), moisture (%) of the paeonia lactiflora root strips, extract (%), paeoniflorin (%) and the like are expressed by average values (x+/-SD); and carrying out statistical analysis on each index.
5.4 test results and analysis
1) Comparison of the weathering characteristics of the respective test lines/varieties
During the 2018-2022 year production trial, the weathered characteristics of each of the test lines/varieties were observed (shown in table 11). The results show that the canal white No. 1 (QBS-1) strain has longer annual growth period, fast emergence and leaf expansion, early flowering and long flowering period compared with the two control varieties ZBSCK and QSCK.
TABLE 11 comparison Table of the characteristics of the candidate lines/varieties of the production test in the main production area of Paeonia lactiflora
Note that: the average of the observation period of each test point of 2018-2022 annual production test is shown in the table.
2) Morphological feature comparison of various reference lines/variety plants in production test
Observations of morphological characteristics of four annual plants of each test point of the test line/variety in the 2018-2022 annual production test show that characteristics of representative plants, leaves, flowers and root systems of the three test lines/varieties (shown in tables 12 and 13) are consistent with morphological characteristics of the multipoint test.
Compared with two control varieties ZBSCK and QSCK, the strain QBS-1 has the outstanding characteristics of high plant size, multiple branches, thick root, crown shape, heavy valve of white flower, and the like, and is consistent with the observation result of a multipoint test. These characteristics lay a good photosynthetic ecological and nutritional ecological foundation for the high quality and high yield of the canal white No. 1 (QBS-1) strain.
Compared with the multipoint test, the biological properties of each test strain/variety and its plants are stable (P > 0.05) in the climatic period within the 5-test-point 2018-2022 year production test; the line QBS-1 has remarkable specificity, stability and consistency.
TABLE 12 statistical table of main morphological characteristics of various reference lines/varieties plants
Note that: the data in the table are the average condition of main morphological characteristics of the strain/variety plants under test at each test point in 2018-2022 production test.
b statistics n=5×3×5.
TABLE 13 characterization of the root bars of the various reference lines/varieties
3) Results of production test and statistical analysis
(1) Production test results
The 2018-2022 year production test shows that the average yield (kg/cell) of each test strain/variety of paeonia lactiflora root trunk is shown in table 14; converted into acre yield (kg/acre) as shown in Table 15.
Tables 14 2018-2022 annual production test yield statistics tables
Note that: the data in the table are the statistical values of the yield of the four-year-old paeonia lactiflora root in autumn harvest of each test point of the production test.
b is the increased yield value of QBS-1 to ZBSCK; c is the QBS-1 over QSCK yield increase value.
Table 15 2018-2022 annual production test conversion mu yield statistics table
Note that: the data in the table are the statistical values of the yield of the four-year-old paeonia lactiflora root in autumn harvest of each test point of the production test.
No significant difference (P > 0.05) in paeonia lactiflora root bar yield for each tested line/variety between 5 trials 3 replicates of the 2018-2022 annual production trial; the peony root strip yields among the 3 test lines/varieties have extremely remarkable difference (P < 0.001), the order of the yields from high to low is QBS-1 (987.67 kg/mu) of Kaubai No. 1, ZBSCK (907.73 kg/mu) of middle Jiang Baihua Paeonia lactiflora, and QSCK (760.07 kg/mu) of mixed populations of Paeonia lactiflora in canal county. The yield of the paeonia lactiflora root stripes of the canal white No. 1 (QBS-1) strain is extremely higher than that of the Jiang Baihua paeonia lactiflora ZBSCK and the QSCK (P < 0.001) of the canal county paeonia lactiflora hybrid population.
The production test and the multi-point test of the second production period have no obvious difference (P > 0.05) between the test points and the repeated paeonia lactiflora root strip yields; however, the paeonia lactiflora root bar yields were lower than in the first cycle multipoint test (2014-2018). The main reason is that the continuous high temperature drought in 2022 leads to the reduction of paeonia lactiflora root yield in the multi-point test and the production test of the second production period (2018-2022).
(2) Analysis of variance and multiple comparisons of production test yields
Analysis of variance of the yields of each of the test lines/cultivars root bars (shown in fig. 23) was performed on the 2018-2022 year production test, and the result showed that the difference in peony yield between 3 test lines/cultivars (f= 12805.14) was significantly greater than the interaction effect of 5 test sites (f=21.11) and test cultivar-test site (f=7.25). The average root yield of the canal white No. 1 (QBS-1) strain is 987.67 kg/mu, and is increased by 8.81% and 29.94% respectively compared with the yield of Jiang Baihua (ZBSCK) root in a control variety (907.73 kg/mu) and the average root yield of the canal county peony hybrid population (QSCK) (760.07 kg/mu), and the yield is increased by 100%.
(3) Stability (mean-coefficient of variation) analysis of production test lines/varieties
The results of analysis of root-strip yield stability (mean-coefficient of variation%) for each of the trial lines/varieties of the 2018-2022 year production test showed that (shown in fig. 24), the canal white No. 1 (QBS-1) line yield stability (cv=0.465%) was greater than the medium Jiang Baihua peony (ZBSCK) control variety (cv=0.588%) and greater than the canal county peony mixed population (QSCK) control variety (cv=1.534%). Indicating stable yield of QBS-1 strain.
(4) Fitness analysis of production test lines/varieties
Analysis results of root-strip yield stability (fitness) of each of the test lines in 2018-2022 production test show that (shown in fig. 25), the fitness of the line of canal white number 1 (QBS-1) and the control variety of medium Jiang Baihua peony (ZBSCK) is 100%.
4) Measurement result and statistical analysis of quality of paeonia lactiflora root strips in production test
(1) The quality measurement results of the Paeonia lactiflora root strips in the production test in 2018-2022 determine the contents of extract and paeoniflorin of the Paeonia lactiflora root strips in each reference strain/variety according to the detection methods and standards of Paeonia lactiflora and Paeonia lactiflora in Chinese pharmacopoeia (2015 edition, 2020 edition, one part). The extract of the root strip of Ganlu No. 1 has an average content of 35.47% and paeoniflorin has an average content of 4.00%. The contents of the extract of the canal white No. 1 strain and paeoniflorin are greatly increased (P < 0.001) compared with those of Jiang Baihua Paeonia lactiflora (ZBSCK) and canal county Paeonia lactiflora mixed population (QSCK) in a control variety. The mass increase point of the canal white No. 1 reaches 100 percent.
Table 16 2018-2022 annual production test drug quality comparison for each of the test lines/varieties (n=3)
Note that: the data in the table are the combined statistical values of the quality indexes of the four-year-old paeonia lactiflora root of each test point of the production test.
(2) Analysis of variance and multiple comparisons of quality of root bars of Paeonia lactiflora in production test
Analysis of variance of the index component content of 3 reference lines/variety root bars at 5 test points in 2018-2022 production test showed that there was a significant difference between extract (%) and paeoniflorin (%) content analysis of variance F values (1388.49, 43.80) between reference lines/varieties as shown in fig. 26. The average content of extract No. 1 (35.47%) and paeoniflorin (4.00%) of the extract are 24.01%, 36.06% and 39.53%, 29.00% (P < 0.001) higher than those of Jiang Baihua Paeonia lactiflora ZBSCK and Paeonia cun county Paeonia lactiflora mixed population QSCK in control variety. The mass increase point of the canal white No. 1 reaches 100 percent.
(3) Stability analysis of quality of Paeonia lactiflora root strips in production test
Quality stability analysis of each of the test lines/variety paeonia lactiflora root bars in 2018-2022 production test (shown in FIG. 27). The results show that the extract stability (mean-coefficient of variation, CV%) of the root strips of different reference lines/varieties of paeonia lactiflora, as well as Jiang Baihua paeonia lactiflora (ZBSCK, cv=0.975%) > canal white number 1 (QBS-1, cv=1.072%) > canal county mixed population of paeonia lactiflora (QSCK, cv=1.666%); indicating that the ZBSCK and QBS-1 extracts are stable. Stability of paeoniflorin (mean-coefficient of variation CV%) was expressed as canal white No. 1 (QBS-1, cv=0.000%) > Jiang Baihua peonies (ZBSCK, cv= 6.369%) > canal county mixed population of peonies (QSCK, cv=10.00%); the content of paeoniflorin in the QBS-1 strain is stable.
(4) Adaptability analysis of quality of paeonia lactiflora root strips in production test
Quality fitness analysis of each of the test lines/variety paeonia lactiflora root bars was performed in 2018-2022 production experiments (fig. 28). The result shows that the adaptability of the extract of the paeonia lactiflora root strip of the QBS-1 strain and the paeoniflorin content at different test points reaches 100%; the adaptability of the paeoniflorin content of Jiang Baihua paeonia (ZBSCK) and the root strips of the canal county paeonia mixed population (QSCK) in 5 test points is 40%; the adaptability of the extract of the root strips of Paeonia lactiflora (ZBSCK) and paeoniflorin content of Jiang Baihua in the culture medium is 0 in 5 test points. The open channel white No. 1 (QBS-1) strain is a good quality peony germplasm resource suitable for Daba mountain.
5.5 production test knots
The observation and statistical analysis results of 2018-2022 annual production test show that the growth and development characteristics of the test strain/variety selected from the main production area of the channel county and the region Jiang Queyao and the biological characteristics, root appearance characteristics, yield and quality of the four annual plants have obvious differences (P < 0.001). Production test the comprehensive evaluation of each of the test lines/varieties is as follows.
1. Line QBS-1
The annual growth period of QBS-1 strain is about 260 days, the seedling emergence and leaf expansion are fast, the plant height is 40-100cm, the average branches of single plant is 15.37+2.38, the root is thick, the flowering period is long, the white flower is heavy, the stamen is petaloid, the carpel number (3-4) is more than 4, and the villus is present. The root yield of the paeonia lactiflora in the production test is 987.67 kg/mu, and the average yield is increased by 8.81% and 29.94% respectively compared with the control varieties ZBCK (Jiang Baihua paeonia lactiflora) and QSCK (mixed peony group in the county); the average content of extract is 35.47%, and the average content of paeoniflorin is 4.00%, which is 24.01%, 36.06%, 39.53% and 29.00% higher than the content of Jiang Baihua paeonia ZBSCK and canal county paeonia hybrid population QSC in the control variety respectively. The QBS-1 strain has stable yield and quality.
2. Control variety of Paeonia lactiflora (ZBSCK) of medium Jiang Baihua
The annual growth period of ZBSCK control variety is about 240 days, the plant height is 40-70cm, the average branches of single plant is 12.75+2.18, the root is thick, the flowering period is longer, the white flowers, the stamen-valve chrysanthemum flowers, the number of carpels (3-4) is 3 more and fluff exists. The root yield of the paeonia lactiflora in the production test is 907.73 kg/mu respectively; the contents of extract and paeoniflorin were 28.60% and 2.87%, respectively. The yield and quality are stable.
3. Canal county peony hybrid population (QSCK) control variety
The annual growth period of QSCK control variety is about 250 days, the plant height is 40-100cm, the average branches of single plant is 11.23+2.81, the root is thicker, the flowering period is longer, white flowers and pink are mixed, the number of carpels is more than 3 (2-4), and the white flowers have villi and pink have no villi. The root yield of the paeonia lactiflora in the production test is 760.07 kg/mu respectively; the contents of extract and paeoniflorin were 26.07% and 3.33%, respectively. The yield and the quality are more stable.
In conclusion, the QBS-1 strain is white peony, has long growing period and flowering period, fast emergence and leaf expansion, high plant height and strong branching force; the root strips are more and thicker, and the root strip yield is high; the extract and paeoniflorin content are high, which accords with the regulations of Chinese pharmacopoeia (2015 edition and 2020 edition, one part). The characteristics of the weather period, biological characters, root appearance characters, yield and quality of each test strain/variety in the production test are consistent with the results of continuous two-production-cycle multipoint tests. There was no significant difference in field performance of disease resistance between QBS-1 line and control line.
6. Comprehensive evaluation of New strains
After the multi-point test and the 2018-2022 production test of two continuous production periods (2014-2018 and 2018-2022) of 3 test lines/varieties at 5 test points, the comprehensive evaluation of plant base and morphological characteristics, fertility characteristics and biological characteristics of a canal white number 1 (QBS-1) line bred by Paeonia lactiflora in a canal county and the yield and quality of Paeonia lactiflora root strips is as follows:
1) The strain QBS-1 is obtained by systematic breeding from traditional Chinese herbaceous peony cultivation mixed population of the town bridge village of eastern Ann and town of the main production area of the white peony road of Sichuan, and the basic source is Chinese herbaceous peony Paeonia lactiflora pall.
2) The annual average growth period of QBS-1 is about 260 days, and the method has the remarkable characteristics of long growth period, fast emergence and leaf expansion, long flowering period, white flower weight, stamen, imperial crown shape and the like. Is a perennial upright herb plant, has thick and strong root and is cylindrical. The plants are high and large, the branching force is strong, and the stem height is 40-100cm. The stem leaves are two-cycle three-out compound leaves; the small blade is in the shape of a needle or an egg, the edge of the small blade is provided with white bone fine teeth, the two sides of the small blade are hairless, and the back of the small blade is thin and short and soft along the veins. The diameter of the top of the peanut stem and the axillary is 8 cm to 13cm; 4-5 bracts, in the shape of a needle and a leaf; sepals 3 or 4, wide oval or nearly circular; the petals are white, the crown shape is a heavy petal, and the petals are inverted oval; a carpel (2-) 4 with nap; the column head is red. The pod does not develop normally and is tightly fluffed. The flowering period is 4-5 months. There was no significant difference in field performance of disease resistance between the canal white No. 1 and the control variety.
3) A multi-point test of two continuous production periods (2014-2022 years) shows that the average yield of the strain of the Paeonia lactiflora root strip (QBS-1) is 1023.17 kg/mu, and the yield is increased by 8.09% and 29.48% compared with the control varieties of the Zhong Jiang Baihua Paeonia lactiflora (ZBSCK) and the Fangxian Paeonia lactiflora mixed population (QSCK) respectively (P < 0.001). Wherein, the average yield of the canal white No. 1 is 1053.53 kg/mu in the first production period (2014-2018 years) multipoint test, and the yield is increased by 7.55% and 28.87% respectively compared with the control varieties of the medium Jiang Baihua Chinese herbaceous peony (ZBSCK) and the canal county Chinese herbaceous peony mixed population (QSCK) (P < 0.001); in the second production cycle (2018-2022 years) multipoint test, the average yield of the canal white No. 1 is 992.80 kg/mu, and the yield is increased by 8.68% and 30.14% respectively (P <0.001 x) compared with the control varieties of the medium Jiang Baihua Chinese herbaceous peony (ZBSCK) and the canal county Chinese herbaceous peony mixed population (QSCK). The yield increase point of the canal white No. 1 reaches 100 percent.
Statistical analysis of paeonia lactiflora root strip yield in 2018-2022 production experiments shows that average yield of the strain of Gansu No. 1 is 987.67 kg/mu, and the average yield is increased by 8.81% and 29.94% respectively (P < 0.001) compared with a control of Chinese Jiang Baihua paeonia lactiflora (ZBSCK) and a control of mixed-population of Paeonia lactiflora (QSCK) in county. The yield increase point of the canal white No. 1 reaches 100 percent. The characteristics of the production test such as the physical period, biological characters, root appearance characters, yield and quality of each test strain/variety are consistent with the statistical analysis results of continuous two-production-period multipoint tests.
A multipoint test of two consecutive production cycles (2014-2018 and 2018-2022) and a production test of 2018-2022, statistical analysis of the contents of effective pharmaceutical component extract and paeoniflorin of each strain/variety of paeonia lactiflora strips showed that: two consecutive production cycle multipoint experiments, average extract (35.73%) and paeoniflorin content (3.97%) of the line of canal white 1 (QBS-1) were upgraded by 24.51%, 36.04% and 32.22%, 11.21% (P < 0.001) respectively compared to the control varieties of medium Jiang Baihua paeonia (ZBSCK), mixed-population of canal county paeonia (QSCK). In 2018-2022 production test, average content of extract of strain of Ganba No. 1 and paeoniflorin (35.47% and 4.00%) is respectively improved by 24.01%, 36.06% and 39.53%, 29.00% (P < 0.001) compared with control varieties of Chinese Jiang Baihua radix Paeoniae (ZBSCK) and mixed population of radix Paeoniae (QSCK) in Qingxian county. The mass increase points of the canal white No. 1 reach 100 percent.
4) The average content of paeonia lactiflora strips in the line QBS-1 (QBS-1) is detected as follows: the water content is 8.6%, the total ash content is 3.2%, sulfur dioxide is 27.89mg/kg, lead is 0.14mg/kg, arsenic is 0.16mg/kg, mercury is 0.003mg/kg, copper is 3.8mg/kg, extract is 35.7%, paeoniflorin is 3.97%. The characteristics and quality of plant-based primordium and root of the canal white No. 1 strain accord with the regulations of white peony root and red peony root in Chinese pharmacopoeia (2020 edition, one part).
In conclusion, the source of the QBS-1 is clear, the genetic character is stable, the field performance is tidy and consistent, and the novel strain is a new strain of paeonia lactiflora, and has obvious difference from a control variety.
7. The technical key points of cultivation
The cultivation technical key points of the strain comparison test, the multipoint test and the production test are as follows:
1) Environmental conditions of the producing area
(1) Climate conditions
The medium and subtropical humid climate zone is suitable for hilly and low mountain ecological zone, the altitude is 200-1000m, the annual average temperature is 14-18 ℃, the annual average sunshine hours are 1100-1200h, the annual rainfall is 800-900mm, the relative humidity is about 80%, and the annual frost-free period is more than or equal to 290 days.
(2) Soil condition
The soil layer thickness of hills and low mountains is more than or equal to 50mm, and the sandy or loam with pH of 6.5-8.5 is suitable for loose, fertile and well-drained soil. Continuous cropping is contraindicated and rotation is carried out for 2-3 years (season).
(3) Ecological environment comparison of major production area of Sichuan white peony root
The production environment should meet the regulations of NY/T5010-2016. The traditional road-region major production area of the Sichuan peony is located in the subtropical humid climate zone in China in Jiang county and canal county. The midrange Jiang Queyao is located in the purple hills or suitable ecological areas of low mountains (base 482m, north latitude 31 deg. 10'33 "N, east longitude 104 deg. 36' 59" E) at an altitude of 400-1000m in northwest Long Quanshan. The production area of Paeonia lactiflora in the county is located in the mountain of Daba, the purple hills of 300-1000m or the suitable ecological area of the low mountain (base 642m, north latitude 30 DEG 56 '15' N, east longitude 107 DEG 09 '51' E). The ecological environment of the producing areas of Zhongjiang and Chen county is suitable for the production of high-quality white paeony root.
2. Cultivation technique
(1) Seed selection
(1) Bud head planting standard: cutting off the bud-bud reed heads of the paeonia lactiflora from the roots when the seedling of the paeonia lactiflora plant is poured, withered and harvested in autumn; and selecting the reed heads with strong buds as propagation materials, wherein the reed heads have no plant diseases and insect pests, rot, hollow and dry shrinkage. Cutting the reed heads into small blocks according to the natural growth characteristics, wherein each block needs to retain -3 robust buds; and 3cm of reed heads are required to be reserved.
(2) Planting and selecting bud head: when the paeonia lactiflora roots are harvested in autumn, bud heads meeting the requirements are selected for planting, and the crude roots below the bud heads for planting are cut off for medicinal use; the bud-bearing reed heads left are used as asexual propagation materials. And (5) planting the bud heads for soil or sand storage. Sand cultivated in the storage bud head is disinfected by 70% thiophanate methyl 1000 times liquid. After disinfection, the water content of the sand is agglomerated by hand, and the sand is released to be dispersed.
(2) Planting technique
(1) Planting time: asexual reproduction and autumn (10 months) cultivation.
(2) Preparing soil and fertilizing: and selecting sandy loam with deep soil layer, good water drainage, looseness and fertility to plant paeonia lactiflora. After the front crop is harvested, sundries such as weeds, stones, front crop residues and the like in the soil are removed; applying 1000-1500kg of decomposed organic fertilizer per mu, deep ploughing soil for more than 40cm, ploughing for multiple times, trimming, raking, and digging drainage ditches around the planted land.
(3) And (3) planting the paeonia lactiflora in a high-box-planting mulching film-covering mode, namely planting the paeonia lactiflora in a high-box planting and water-saving grass-preventing mulching film-covering planting mode for 10 months. The height of the carriage is 30-35cm, and the width of the inter-carriage ditch is 20-30cm. After planting and covering soil on the head of the field planting paeonia lactiflora, covering high-quality black mulching films. Stretching and paving the black film, and covering the black film tightly on the carriage surface; the periphery of the edge of the membrane is compacted and sealed by fine soil. The planting specification is 30cmx 40cm, and the blunt pry is used for opening the holes in a T shape; applying 1500-2000kg of organic fertilizer per mu. After the fertilizer is permeated and dried under water, the bud heads planted by the peony bud heads are upwards pressed into the nest, and fine soil is covered for 3-6cm.
(3) Field management technology
(1) Checking for missing and supplementing seedlings: after the seedlings emerge in spring, if the field seedlings are found to be lack, the seedlings are timely reseeded so as to ensure the seedling alignment.
(2) And (3) cultivating and weeding: in the first year and the second year, the seedlings are small, the roots of the white peony are fine, the roots are not deep, the weeds are easy to grow in the soil, the soil is loosened by shallow intertillage, the weeds in the field are removed in time, and the seedling is carried out 2-3 times per year in combination with fertilization.
Removing weeds which grow in autumn and winter after spring and before emergence of seedlings for the first time, and shallow intertillage and soil loosening; secondly, when weeds are bred in a large amount in summer, the weeds are removed, and shallow intertillage loosens soil without damaging young roots; after the seedlings are fallen in autumn for the third time, the weeds are removed, sundries such as dead branches, fallen leaves and the like in the field are cleaned, and the seedlings are transported to the outside of the field for centralized treatment.
In the third and fourth years, the intertillage weeding is carried out for 1-2 times in early spring and early summer.
(3) Timely topdressing: after autumn planting, topdressing starts in the next year.
Topdressing 4 times in the first year. Combining cultivation and weeding for the first time in 3 months, and applying 1500-2000kg of decomposed organic fertilizer per mu; applying 1500-2000kg of organic fertilizer and 20kg of decomposed cake fertilizer for each mu in the growing period of 4 months and 5 months respectively for the second time and the third time; the fourth time is 11 months, 2000kg of organic fertilizer is applied per mu.
Topdressing for 3 times in the second and third years. 1500kg of decomposed organic fertilizer and 20-30kg of biological fertilizer are applied to each mu of the fertilizer for the first time in 3 months. And applying 1500kg of decomposed organic fertilizer and 25kg of decomposed cake fertilizer per mu in 4 months for the second time. And applying 2000kg of decomposed organic fertilizer and 30kg of calcium superphosphate per mu in 11 months for the third time.
Topdressing is carried out in the fourth year, and 2 times of topdressing are carried out before harvesting. The biological fertilizer is applied for 25-30kg, the decomposed cake fertilizer is 40-50kg and the farmyard manure is 1500kg in 2 months for the first time. And applying 1500kg of farmyard manure and 25kg of decomposed cake manure to each mu in 4 months for the second time.
(4) Picking out flower bolts: when the paeonia lactiflora plants are bolting and bud emergence in spring every year, the buds are removed in time.
(5) Drought-resistant and water-logging-draining: after the paeonia lactiflora is planted and survived, if drought occurs, the paeonia lactiflora should be irrigated in time to resist drought and protect seedlings. 7-9 months into rainy season, summer heavy rain and autumn soft rain easily cause excessive wetting of soil, cause soil ponding root rot and seedling death, drain water in time and prevent waterlogging, and four side ditches and middle ditches of the peony planting land are prepared, and the cage ditches are drawn out in time.
(4) Comprehensive pest control technology
The diseases and insect pests of Paeonia lactiflora mainly comprise root rot, powdery mildew, gray mold, round spot, rust, grubs, cutworm and the like.
(1) Agricultural control: selecting a land block with high topography, good ventilation and loose soil; selecting healthy and strong white peony heads without diseases, soaking seeds; deeply turning kang soil; reasonably rotation, close planting and fertilization; strengthening water and fertilizer management; timely cultivating and weeding, removing old leaves of disease plants and disease of plant bases, and cleaning a field.
(2) Physical control: and (3) installing frequency vibration type insecticidal lamps and the like on the concentrated planting places of the paeonia lactiflora to trap and kill adults of underground pests.
(3) Chemical control: pesticide application should meet DB51/337 requirements. The main diseases and insect pests and recommended control methods are shown in Table 17.
Table 17 main diseases and insect pests of paeonia lactiflora and recommended prevention and control method table
3) Harvesting and post-harvesting treatment technology
(1) Harvesting and planting Paeonia lactiflora in proper period for 4 years, withering Paeonia lactiflora in autumn (9-10 months), and selecting for sunny days. Firstly cutting off stems and leaves, then digging out all the paeonia lactiflora root strips, removing soil, and transporting the paeonia lactiflora root strips into a room.
The new line of the Yuan Bai No. 1 is collected after 10 months of peony plants wither; harvesting the single plants and the plant line comparison test autumn seeds after 3 years; strain comparison test, multipoint test, production test, and harvesting after autumn seed of 4 years.
(2) Post-mining treatment
Cutting off root strips of Paeonia lactiflora from the place where Paeonia lactiflora heads grow, cutting off side thin roots on thick roots, trimming the convex surface, cutting off heads and tails, and stacking in a room according to large, medium and small sizes.
(3) Processing in producing area
Decocting radix Paeoniae alba: the Chinese herbaceous peony root strips are graded according to the thickness, and are respectively boiled by water, wherein the water quantity is suitable for submerging the Chinese herbaceous peony root strips, and the boiling is suitable for thoroughly boiling. The boiling time depends on the thickness of the root strip of Paeonia lactiflora, so it is preferable that the root strip of Paeonia lactiflora has no white heart.
Peeling: taking out the boiled radix Paeoniae strips from the pan, immediately washing with cold water, removing the outer skin of the strips with a machine, scraping with a bamboo knife, and removing insect eyes.
And (3) drying: conveying peeled radix Paeoniae to sunning ground in time, spreading, and drying for 1-2 hr, and piling radix Paeoniae strips for thick drying. The utility model is turned up and down every day. Covering radix Paeoniae alba with bamboo mat or the like when sun is too violent in noon; and (5) spreading out and airing after 15:00-16:00 hours. Repeating for 3-5 days, stacking "sweating" in room for 2-3 days, and air-drying for 3-5 days. In the processing process, if the weather is long rain, the paeonia lactiflora root strips are baked by slow fire.
(4) The quality requirements are as follows: meets the regulations of Chinese pharmacopoeia (2015 edition before breeding and 2020 edition after breeding, one part).
4) Packaging and storage
(1) And (3) packaging: the packaging material meets the requirements of GB/T6543 and GB 9693.
(2) And (3) storage: the mixture is stored in a clean, dry, cool, ventilated and odorless warehouse, the temperature is controlled below 25 ℃, and the relative humidity is less than or equal to 70 percent.
8. Identification of primordia
On 28 th year of 2022, the medical science and technology is invited to take the plant form, flower characteristics, root form, color, section characteristics, smell and other characters and identify for the canal white No. 1 (QBS-1) by inviting the university of traffic Chinese medicine institute of Sichuan province Li Yuecheng to teach to go to the canal county peony-road-area of production Zhang Xingguo. Through field observation and picture identification, the conclusion is that: the strain QBS-1 is Pall of Paeonia lactiflora Paeonia lactiflora belonging to Ranunculaceae, and meets the requirements of Paeonia lactiflora and Paeonia lactiflora in Chinese pharmacopoeia (2020 edition, one part).
9. Stamen flap gene assay
The AG and MADS2 genes associated with stamen of ZBSCK peony-lobed stamen of Fangzu No. 1 (QBS-1) and of Paeonia lactiflora in Jiang Baihua were determined by qRT-PCR using light purple pink-lobed stamen as a control. The results showed that the average expression level of AG gene in the canal white No. 1 lobed stamens (23061.67, n=3) was only 51.6% of the average expression level in the middle Jiang Baihua peony lobed stamens (44705.56, n=6), while the average expression level of MADS2 gene in the canal white No. 1 lobed stamens (11.532, n=3) was 3.3 times of the average expression level in the middle Jiang Baihua peony lobed stamens (3.5, n=6). The genes of the Paeonia suffruticosa petal stamen of the open channel white No. 1 are different from those of the Paeonia suffruticosa of the Medium Jiang Baihua.
10. Evaluation of novel System characteristic Properties
1) Morphological characteristics of plants
The strain QBS-1 is perennial upright herb, with thick and strong root and cylindrical shape. The plant is big, branches are more, and the stem height is 40-100cm. The stem leaves are two-cycle three-out compound leaves; the small blade is in the shape of a needle or an egg, the edge of the small blade is provided with white bone fine teeth, the two sides of the small blade are hairless, and the back of the small blade is thin and short and soft along the veins. The diameter of the top of the peanut stem and the axillary is 8 cm to 13cm; 4-5 bracts, in the shape of a needle and a leaf; sepals 3 or 4, wide oval or nearly circular; the petals are white, the crown shape is a heavy petal, and the petals are inverted oval; 3-4 parts of carpel with villus; the column head is red. No mature fruit. The flowering period is 4-5 months.
2) Growth and development characteristics
The strain QBS-1 has the remarkable characteristics of long growth period, fast emergence and leaf expansion, strong branching force, long flowering period and the like, and has the annual average growth period of about 260 days. The characteristics lay a good photosynthetic ecological and nutritional ecological foundation for the high quality and high yield of the canal white No. 1 strain.
The multipoint test and production test research show that the biological properties of each test strain/variety and plants thereof are stable (P > 0.05) in the climatic period of the continuous two production cycles multipoint test at 5 test points. The canal white No. 1 strain bred from the canal county paeonia district main production area system has obvious specificity, stability and consistency. Disease resistance of the canal white No. 1 and two control varieties is not obviously different in field performance.
3) Yield characteristics
In 2014-2022 continuous two-production-period multipoint test, the average mu yield of the root trunk product of the strain of Gande No. 1 is 1023.17kg, and the yield is increased by 8.09% and 29.48% respectively (P < 0.001) compared with the control varieties of the mixed group QSCK of Chinese herbaceous peony ZBSCK and Gandelia and Paeonia in China. In 2018-2022 production test, average mu yield of the white ditch No. 1 is 987.67kg, and the yield is increased by 8.81% and 29.94% respectively compared with the two control varieties (P < 0.001). The yield increasing point reaches 100 percent. Statistical analysis results of the physical stage, biological properties, root strip appearance properties and yield characteristics of each test strain/variety in the multi-point test and the production test are consistent.
4) Quality characteristics
Average content of root bars of the canal white number 1 (QBS-1) strain: 35.7% of extract, 3.97% of paeoniflorin, 8.6% of water, 3.2% of total ash, 27.89mg/kg of sulfur dioxide, 0.14mg/kg of lead, 0.16mg/kg of arsenic, 0.07mg/kg of mercury, and 3.8mg/kg of copper. The quality of the canal white No. 1 accords with the regulations of Chinese pharmacopoeia (2015 edition, 2020 edition, one part).
The foregoing description of the embodiments has been provided for the purpose of illustrating the general principles of the invention, and is not meant to limit the scope of the invention, but to limit the invention to the particular embodiments, and any modifications, equivalents, improvements, etc. that fall within the spirit and principles of the invention are intended to be included within the scope of the invention.
Claims (10)
1. The breeding method of the new peony strain is characterized by comprising the following steps:
s1, establishing a peony germplasm resource database: establishing a peony germplasm resource nursery and a peony excellent germplasm resource expansion breeding nursery, collecting peony nursery germplasm data, and establishing a peony germplasm resource database;
s2 strain comparison test: screening a plurality of excellent variant single plants from a database, carrying out a plurality of period strain comparison tests on the excellent variant single plants and a local excellent hybrid population, observing the weather characteristics, biological properties, yield, quality and resistance of the single plants, screening a plurality of excellent strains, and purifying the excellent strains to form stable excellent strains;
s3 strain comparison test: carrying out a plurality of cycles of strain comparison tests on stable good strains and local good hybrid populations, observing the climatic period characteristics, biological characters, yield, quality and resistance of the single strain, and screening out new peony strains;
s4 multipoint test: carrying out multi-point tests for a plurality of periods on the new paeonia lactiflora strain, the local hybrid group good strain and the foreign good strain, observing and comparing the weather characteristics, biological properties, yield, quality and resistance of each tested strain/variety, and verifying the yield, specificity, stability and consistency of the new paeonia lactiflora strain;
S5 production test: performing production tests on the new paeonia lactiflora strain, the local hybrid group good strain and the foreign good strain, observing and comparing the characteristics of the weathers, the biological properties, the yield, the quality and the resistance of each tested strain/variety, and verifying the yield, the specificity, the stability and the consistency of the new paeonia lactiflora strain;
s6, field technology identification: performing primitive identification on the new strain of paeonia lactiflora, and identifying the characteristics of plant morphology, flower characteristics, root morphology, color, section characteristics, smell and the like of the new strain; determining AG and MADS2 genes related to stamen petal in paeonia lactiflora new strain and foreign excellent strain by qRT-PCR method;
s7, evaluating the characteristic characteristics of the new strain: and comprehensively evaluating the plant base and morphological characteristics, the fertility characteristics and the biological characteristics of the new variety and the yield and quality of the paeonia lactiflora root bars, wherein the comprehensive evaluation content comprises the plant base and morphological characteristics, the fertility characteristics and the biological characteristics and the yield and quality of the paeonia lactiflora root bars.
2. The breeding method of new strain of paeonia as claimed in claim 1, wherein the biological characteristics and the biological yield of the aerial part and the underground part of the plant are respectively measured in the S4 multi-point test step and the S5 production test step.
3. The method for breeding new strain of paeonia as claimed in claim 2, wherein, when determining biological characteristics and biological yield, the yield of the dry product cell of the test strain/variety of four-year-old paeonia is converted into mu yield, and then statistical, variance and multiple analysis are performed, including single analysis and multi-cycle combination analysis of each cycle.
4. The breeding method of new strain of paeonia as claimed in claim 1, wherein, in the step of S4 multipoint test and the step of S5 production test, when the quality of the root of the four-year old paeonia of the reference strain/variety is observed and compared, the measured indexes are extract (%) and paeoniflorin (%), and then statistical and variance and multiple analysis are performed, including single analysis of each period and multi-period combination analysis.
5. The breeding method of a new strain of paeonia lactiflora in accordance with claim 3 or 4, further comprising stability analysis and fitness analysis.
6. The breeding method of new peony strain according to claim 1, wherein in the step of S4 multipoint test and the step of S5 production test, sandy loam with uniform fertility, deep soil layer, flat soil and good drainage is selected experimentally.
7. The breeding method of new peony strain according to claim 1, wherein in the step of S4 multipoint test and the step of S5 production test, protection rows are arranged on two sides of a field cell, and a local hybrid group excellent strain is planted.
8. The breeding method of new strain of paeonia as claimed in claim 1, wherein, in the step of S4 multipoint test and the step of S5 production test, the observed data of morphological feature values of plants when biological traits are observed comprises: plant height, branch number, flower color, petal, carpel number, plush and leaf characteristics, and root strip appearance characteristic observation data comprise: skin color, section color, presence or absence of wood core and root bar features.
9. The breeding method of new strain of paeonia as claimed in claim 1, wherein in the step of S2 strain comparison test, the row spacing of the planted strain is 30cm multiplied by 40cm, the planted strain is cultivated in autumn, and the planted strain is harvested in autumn after 3 years and randomly arranged; in the S3 strain comparison test step, the row spacing of the planted strains is 30cm multiplied by 40cm, the cell specification is 1.6mmultiplied by 1.5m=2.4 square meters, 5 strains/row multiplied by 4 rows=20 strains are planted in each cell, and the plants are cultivated in autumn, harvested in autumn in 4 years and randomly arranged; in the S4 multipoint test step, the field cells are designed to be 3 multiplied by 3, the field cells are arranged in random groups, the row spacing of the planted plants is 30cm multiplied by 40cm, the specification of the cells is 1.6mX3.0m=4.8 square meters, and 10 plants/row multiplied by 4 rows=40 plants of the planted peonies are arranged in the cells; in the S5 production test, the field cell is designed to be 3 multiplied by 3, the random group arrangement is adopted, the row spacing of the planted plants is 30cm multiplied by 40cm, the cell specification is 4.0mX6.0m=24 square meters, and 20 plants/row multiplied by 10 rows=200 plants of the cell planted Chinese herbaceous peony.
10. The method for breeding new strain of paeonia as claimed in claim 1, wherein in the steps of the S2 strain comparison test, the S3 strain comparison test, the S4 multipoint test and the S5 production test, cultivation considerations of paeonia include: the method comprises the steps of producing environment conditions, a cultivation method, a disease and insect comprehensive control method, a harvesting and post-harvesting treatment method, packaging and storage.
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