Cultivation Technology and Research Advance on the Germplasm Resources of Angelica dahurica

Rong SONG Bei YAN Qiujiao WANG Jiamin ZHOU Xiaoqi ZHU

Abstract The roots of Angelica dahurica are a commonly used traditional Chinese herbal medicine. It has the effects of dispelling wind and relieving pain， expelling pus and dispersing swelling， clearing the nasal orifices， and eliminating dampness. In recent years， the cultivation of A. dahurica in Hunan Province has developed rapidly， but the research on new varieties selection and standardized cultivation techniques is lagging behind. This paper reviewed the germplasm resources， variety distribution and standardized cultivation technology of A. dahurica in China， hoping to provide technical support for the development of A. dahurica industry.

Key words Angelica dahurica; Germplasm resources; Cultivation technologies

The Chinese Pharmacopoeia （2015 edition） stipulates that the Chinese medicinal material Angelica dahurica is dry roots of A. dahurica （Fisch. ex Hoffm.） Benth. et Hook. F. or A. dahurica （Fisch. ex Hoffm.） Benth. et Hook. f. var. formosana （Boiss.） Shan et Yuan. A. dahurica is a traditional Chinese herbal medicine with pungent taste and warm nature. It has the effects of dispelling wind and relieving pain， expelling pus and dispersing swelling， clearing the nasal orifices， and eliminating dampness. It is mainly used to treat common cold due to wind-cold， headache， rhinitis， toothache， leukorrhea with reddish discharge， and sore swelling and pain[1-2]. The chemical components of A. dahurica plant include coumarin， volatile oil， phytosterols， alkaloids， polysaccharides， mineral elements， etc. The main medicinal components are coumarin and volatile oil[3]. With the development of modern medical technology， the chemical composition and main medicinal components of A. dahurica have been more clearly defined. Its anti-inflammatory， antibacterial， antipyretic and analgesic， antispasmodic， anticancer， antiradiation， cardiovascular， and photosensitivity effects have also been discovered. As a result， the market demand for A. dahurica has increased， and the development of artificial planting area has accelerated. In addition to its medicinal use， it has also been widely used as an industrial raw material in the fields of fragrances， cosmetics and edible fragrances in daily chemical products.

Hunan Province was once the dominant producing area of A. dahurica. Chaling County has a long history of A. dahurica cultivation， and the cultured A. dahurica is well-known throughout the country for its high content of coumarin and volatile oil. Chaling A. dahurica is chrysanthemum-like in appearance， large in size， white in color， and has no tendons. It has strong fragrance and is the top grade among A. dahurica. Chaling A. dahurica was called "Chuzhi" in ancient times， and was once listed as one of the three national famous A. dahurica materials with "Hangzhi" and "Chuanzhi". In the Encyclopedia of National Local Products published in 1987， Chaling A. dahurica was ranked first on the list of A. dahurica and known as one of the "Three Treasures" of Chaling. Since the 1990s， with the decline in the price of A. dahurica， the peoples enthusiasm for planting has declined sharply， and the planting area of A. dahurica in Hunan has shrunk sharply. In recent years， with the rising market of A. dahurica， the planting area of A. dahurica in Hunan has increased year by year. This paper summarized the research situation of A. dahurica in recent years from germplasm resources， production environment and cultivation technology of A. dahurica， aiming to provide references for the comprehensive utilization of the traditional Chinese medicine A. dahurica in Hunan.

Germplasm Resources

Germplasm resources， also called genetic resources， include all available genetic materials such as wild species， wild relatives and cultivated species. The germplasm resources of medicinal plants are the source of the production of Chinese medicinal materials， and are the basis for evaluating， using， and preserving different medicinal plant varieties （lines）[4]. Their quality plays a decisive role in the quality and yield of Chinese medicinal materials[5]. Investigating， sorting out and comprehensively evaluating the germplasm resources of A. dahurica from different origins is of great significance for the protection and utilization of germplasm resources of A. dahurica. Screening of excellent strains is of great significance for ensuring uniform and stable quality of medicinal materials.

Wild germplasms

The wild germplasm source of the traditional Chinese medicine A. dahurica should be the relative species of A. dahurica， including the original variant Xingan Baizhi and the variety Taiwan Baizhi. There have always been controversies about the original plant and classification of A. dahurica， and there is no consensus yet. Most scholars believe that Xingan Baizhi is not a wild species of A. dahurica， but a relative species. There are also studies believing that the wild species of A. dahurica is Taiwan Baizhi. Some scholars speculate that there may be Taiwan Baizhi in Taiwan， Fujian and Zhejiang coastal areas， but it has not been found yet. Xingan Baizhi， also known as Hebei Duhuo， Dahuo， Xiangdahuo， Zoumaqin， Zoumaqin Tongzi and Langshanqin， is widely distributed， and grows in a large quantity in the wild in the Northeast. It distributes between 300-370 m above sea level， grows mostly in moist grasses， shrubs， and sandy soil near streams， and has not been cultivated artificially yet[6].

Cultivated varieties

Traditional commercial medicinal materials of A. dahurica include Hang Baizhi， Chuan Baizhi， Yu Baizhi and Qi Baizhi， which are mainly produced in Zhejiang， Sichuan， Henan and Hebei， and are all cultivated varieties. In recent years， with the development of the A. dahurica industry and the strong adaptability of A. dahurica， it has been introduced and cultivated in many areas， and have formed a certain scale in some areas which become the new main producing area of A. dahurica， such as Bozhou， Anhui[7].

Chuan Baizhi is a famous authentic medicinal material produced in Sichuan， which accounts for about 70% of the countrys commercial A. dahurica production[8]. The original plant of Chuan Baizhi is Hang Baizhi， which is mainly produced in Daxian County， Suining， Qu County， Anyue， Nanchong and other places of Sichuan. A. dahurica produced in Chongqing is mainly cultivated in Sanquan Town， Nanchuan District. Suining is known as the "Hometown of A. dahurica in China". It is located in the middle of the Sichuan Basin and has many years of planting history. Its soil and climate are very suitable for the growth of A. dahurica. In 2006， Suining passed the GAP base certification of Chuan Baizhi by the State Food and Drug Administration. Its products are large in volume and high in quality， and are sold well in domestic and foreign markets. The A. dahurica produced in Nanchuan production area of Chongqing was introduced in Suining， Sichuan. It has many years of planting history. The sowing and collection of medicinal materials and seeds of A. dahurica in Sanquan Town are all later than other Chuan Baizhi production areas. The towns annual output once reached 2 500 t. At present， due to continuous cropping all the year round， the land is becoming barren， pests and diseases have increased， and the yield has dropped significantly[6].

Hang Baizhi is mainly produced in Jiangsu and Zhejiang， mainly grown in Yuyao， Jianqiao， Linhai， Hangzhou， Yuhang and other places. In recent years， due to urban development， arable land has shrunk and output has been decreasing year by year. Its main production areas have moved to Dongyang and Panan. The planting area is also very small. In these producing areas， A. dahurica is generally self-produced and sold， and bulk products are generally Chuan Baizhi.

Yu Baizhi is mainly produced in Henan Province and is the customary name for A. dahurica produced in Yuzhou and surrounding counties and cities in Henan. The banks of Shengtai River and Xiaohong River in Changge County are also one of the main producing areas of Yu Baizhi. Yu Baizhi was once exported to Southeast Asia， but is now mainly used in northern provinces.

Qi Baizhi is mainly produced in Anguo， Hebei. In the early days of liberation， the yield of Qi Baizhi was only 1 125-1 500 kg/hm2. In the 1980s， the yield was 6 000 kg/hm2 with reasonable dense planting. In the past few years， the output of Qi Baizhi was relatively small. The vast majority of A. dahurica in the Anguo medicinal materials market is Chuan Baizhi， and a small part is Qi Baizhi. Pharmacists generally believe that Qi Baizhi is "big head， hollow center， and poor quality"， and Chuan Baizhi is of high quality and has a price higher than Qi Baizhi. In recent years， due to the increase in the price of the whole commodity A. dahurica， and the planting area of A. dahurica has increased[9].

Bo Baizhi has a short cultivation history. It was introduced from Yuzhou of Henan， Anguo of Hebei and Zhejiang in the 1970s. Bo Baizhi is mainly cultivated in Shijiuli Township， Dasi Township， Shibali Town and Qiaodong Town of Bozhou[10]. It has developed rapidly in recent years and its cultivation area is relatively large. However， Bo Baizhi is mainly used for spices and sold to the wholesale market of spices in Tengzhou City， Shandong Province. The raw materials are sliced for sale and wholesale to all parts of the China. Heze of Shandong Province is close to Bozhou， Anhui， and in recent years， the cultivation of A. dahurica in Heze has formed a certain scale. It is said that most of the A. dahurica cultivated in Heze was introduced from Bozhou， which is a series with Bo Baizhi.

Genetic relationship between germplasms

Cytological analysis

Wang et al.[11] did a cytological analysis of different germplasms of A. dahurica and believed that the traditional Chinese medicine A. dahurica was closely related to Xingan Baizhi and Taiwan Baizhi; and 4 cultivated samples （Chuan Baizhi， Hang Baizhi， Qi Baizhi， Yu Baizhi） were very similar in the karyotype of the chromosomes， which proves that there is no difference in cell taxonomy， and the karyotype of A. dahurica in Angelica plants is a relatively unique type. The karyotype formula is 2n=22=12m+2msat+4sm+4st; the 1st to 7th pairs are central centromere chromosomes， the 8th and 9th pairs are subcentral centromere chromosomes， and the 10th and 11th pairs are proximal centromere chromosomes; the length of chromosomes is 6.7-10.9 μm， the total chromosome length of a single set of chromosomes is 100 μm， and the karyotype asymmetry coefficient is 63.6%， which is slightly asymmetric; and the degree of asymmetry is classified as type 2A.

Molecular biology analysis

In the long-term natural and artificial selection process， medicinal plants adapt to the natural and artificial ecological environment of the producing areas. Under different ecological conditions， the genes of the same plant species or varieties that regulate plant growth and development and secondary metabolic activity will be differentiated， which is manifested by changes in plant morphology and or differences in effective ingredients， which ultimately results in differences in the yield and quality of medicinal materials[12]. After analyzing the karyotype of A. dahurica， Pan et al.[13] found that Chuan Baizhi， Hang Baizhi， Qi Baizhi and Yu Baizhi belong to the same group. Huang et al.[14] analyzed the genomic DNA of more than 17 individuals in 4 populations of Xingan Baizhi， Qi Baizhi and Hang Baizhi using random amplified polymorphic DNA technology （RAPD） and found that Qi Baizhi and Hang Baizhi belonged to the same population and had certain differences from Xingan Baizhi. Yang et al.[15] analyzed Qi Baizhi， Yu Baizhi， Hang Baizhi， Chuan Baizhi， Xingan Baizhi， and Taiwan Baizhi combining molecular genetic analysis and morphological， anatomical and chemical methods. They believed that commercial Baizhi can be divided into two categories， namely Qi Baizhi and Yu Baizhi; the original plant of commercial A. dahurica should be Taiwan Baizhi， and it belongs to the same group as Taiwan Baizhi. ITS sequence determination showed that commercial A. dahurica and Taiwan Baizhi had exactly the same sequence[15]. Guo et al.[7] used ISSR molecular marker technology to analyze the genetic diversity of the four major categories of commercial A. dahurica and Xingan Baizhi in 20 populations. The genetic differences between the cultivated A. dahurica in different producing areas were small， while Xingan Baizhi had genetic differences from the 4 major commercial A. dahurica.

Agricultural Biotechnology2020

Cultivation Techniques

Conditions of producing areas

A. dahurica has strong adaptability， is preferable to a warm， humid， sunny climate， fear of high temperature， drought-tolerant and cold-resistant， and often grows under forests， at forest edges， streams， shrubs and valleys. It has no high requirement for altitude， and can be planted between 50-500 m above sea level. For the planting of A. dahurica， it is advisable to choose sandy loam with flat terrain， fertile soil， deep soil layer， loose texture， and good drainage. It is not suitable for planting on clayey soil， saline soil and low-lying aquatic land. A. dahurica does not require high water content， and is resistant to drought and susceptible to waterlogging. Fields with convenient drainage should be selected. It requires the atmosphere without industrial waste gas emissions nearby. According to the requirements of Chinese herbal medicine GAP， the air environment quality should meet the GB3095-1996 Ambient Air Quality Standards; the irrigation water quality should meet the GB5084-2005 Standards for Irrigation Water Quality; and the soil environmental quality should meet the GB15618-1995 Soil Environmental Quality Standard.

Site selection and soil preparation

Continuous cropping of A. dahurica should be avoided. It is better to choose the land previously planted with gramineous crops or vegetables for field planting. Before soil preparation， decomposed farmyard manure and superphosphate are spread at rates of 30 000-45 000 kg/hm2 and 375 kg/hm2， respectively， and the field is then plowed （25-30） cm， raked and leveled. When making borders， high borders should be made for flat land， and the borders are （30-40） cm high， and （110-130） cm wide. Drainage ditches （30-40） cm in depth should be formed around flat land. Flat borders can be made for slopes[16-18].

Sowing

Sowing time

A. dahurica can be planted in spring and autumn， and autumn sowing is generally adopted in production. Autumn planting is generally started from early September to late October， and can be postponed to early November in warm southern regions. In northern regions with colder climates， it is better to plant before frost. The sowing of A. dahurica should be timely based on the local climate， not too early or too late. Under the condition of sowing too early and growing too vigorously in the very year， most plants bolt and bloom in the second year， which make the roots hollow and rotted and could not be used for medicinal purposes. However， if the sowing is carried out too late， the seedlings will be prone to freezing damage after their emergence. Generally， it is appropriate to sow from late August to early October.

Sowing methods

Before sowing the A. dahurica seeds， the seed membrane should be removed， and then the seed are kept in warm water at 45 ℃ for 10 min， naturally cooled and soaked for 6 h. The seeds are then fished out and dried before sowing. The seeding of A. dahurica can be performed by direct seeding and seedling transplanting， of which direct seeding is suitable in production. Seedling transplanting can easily lead to multiple branches of the main root， poor rhizome growth， and poor appearance and quality. The direct seeding can adopt seeding in drill or hole sowing. Shallow trenches about 10 cm in depth are formed according to the row spacing （20-30） cm for seeding in drill， while holes are formed according to the plant spacing （15-20） cm×row spacing 30 cm in hole sowing. The seeds are spread uniformly， and covered with a thin layer of soil （1.5-2.0） cm， followed by watering thoroughly and keeping the soil moist. The amount of seeds for seeding in drill is about 30 kg/hm2， and the amount of seeds for hole sowing is about 22.5 kg/hm2[16-18].

Field management

Thinning

When the seedlings return to green in the spring and the height is 6 cm， thinning is carried out to remove weak seedlings， and 1 to 2 strong seedlings are left in each hole; and 1 seedling is reserved every 10 cm for seeding in drill.

Cultivating and weeding

During the whole planting cycle of A. dahurica， cultivating and weeding are performed 3-4 times. After sowing A. dahurica in the first year and before emergence， weeds should be removed for 1-2 times. After emergence， weeding should be carried out in combination with thinning. Weeds should be removed by hands. When the soil is hardened， the soil is lightly hoed and loosened about 3 cm. Loosening the soil should not be too deep， otherwise the main roots will not be deep downwards， and there will be more fork roots， which will affect the quality. After the final singling， before the ridges of the plants are closed， when there are too many weeds， cultivating and weeding can be carried out according to the moisture content. Cultivating and weeding should not be performed after ridge closing.

Nutrient and water management

For water management， when drought occurs and moisture condition is not good， watering should be performed in time to keep the soil moist. The ditches must be cleared before the rainy season to ensure smooth drainage and prevent accumulated water from causing diseases.

In terms of top dressing， the top dressing of A. dahurica seedlings should be less and light in the very year of sowing. Before the spring of the next year， the supply of fertilizer and water should be strictly controlled to prevent excessive growth and bolting. If there is a plant bolting and blooming， it should be pulled out in time. The plants are top-dressed （1-2） times before ridge closing， combining with thinning and cultivating. The top dressing uses decomposed cake fertilizer 2 250-3 000 kg/hm2， which can also be replaced by ternary compound fertilizer and human and animal manure， by applying in shallow furrows.

Disease and pest control

Common diseases of A. dahurica include spot blight， root rot， gray spot， etc. The main pests include red spider and aphids. The control measures include agricultural control， physical control， and chemical control.

Agricultural control

Plants with good disease resistance in the field are selected to reserve seeds. Diseased plants and litter in the field should be timely cleaned up. The observation of field growth should be strengthened to timely and accurately predict and prevent diseases. It should be planted in rotation with gramineous crops for more than 2 years.

Physical control

According to the different nature of the pests， from April to July， frequency-vibration insecticidal lamps are installed or yellow sticky insect boards are hung in the field of A. dahurica.

Frequency-vibration insecticidal lamp installation： The lamps are installed at a density of one lamp per 1 hm2， with a distance between the lamps of （80-100） m， and a height of （1.5-1.8） m from the ground. The lamps are distributed in a checkerboard pattern， and the hanging time is from early April to late July. The lamps should not been turned on in a thunderstorm.

For yellow sticky board， we can choose 20 cm×25 cm or 25 cm×30 cm yellow boards. The hanging quantity is determined according to the purpose. When monitoring， 150-300 boards/hm2 are hung， and 600-750 boards/hm2 are hung during control. As to the hanging height， the lower end of the yellow boards should be level or slightly higher than the top of the A. dahurica plants， and the boards are hung with the surface towards the east-west direction.

Chemical control

Spot blight： The spot blight of A. dahurica mainly damages the leaves， causing the leaves to die. In severe cases， it causes plant death， resulting in low quality of A. dahurica medicinal materials and loss of yield of A. dahurica. The control method can be paying attention to clearing the garden after harvest and burning the diseased tissues. In the early stage of the disease， 1∶1∶100 Bordeaux mixture or carbendazim， thiophanate methyl， mancozeb and chlorothalonil can be sprayed[19-21].

Root rot： The root rot of A. dahurica is one of the important diseases in production. It mainly occurs in the processing and drying process after harvest. The general incidence is about 15%， and it can reach 30% in severe cases， or all plants rot. Therefore， attention should be paid to protect the periderm of A. dahurica from being damaged during digging， transportation and fresh storage， and it should be processed and dried in time[19-21].

Black spot disease： The black spot disease of A. dahurica mainly damages the leaves. The pathogen can spread through the summer and winter in the field， and it is easy to spread during the hot and humid seasons， causing serious diseases and affecting yield and quality. Therefore， we should clear the garden well in production to reduce the occurrence of diseases， strengthen field observation， and timely use drugs for sterilization and control when diseases are found[19-21].

Red spider： Red spider is also called Tetranychus cinnabarinus. Adults and nymphs draw their nets and damage the leaves of A. dahurica. Red spiders mainly live through the winter in groups of females on branches， stubbles and the roots of vegetable plants. The overwintering adults go out in March of the following year， with generational overlap. Therefore， it is recommended to clean up the garden during soil preparation and in winter to kill insects and eggs; it is necessary to observe the disease in the seasons of drought in spring and high temperature and less rain in summer， and use imidacloprid and fendicarb to control the disease in the early stage of disease， once a week， 2-3 times in succession[19-21].

Aphids： Aphids mainly damage the young leaves and tops of A. dahurica with adults and nymphs. It is necessary to clear the garden in winter， deep bury or burn the litter， and spray 50% pyrex 1 000-2 000 times liquid， or 40% dimethoate EC 1 500-2 000 times liquid when the pest occurs， once every 7 d， several times in succession[19-21].

Harvesting and processing

From July to August of the following year， the leaves of A. dahurica can be harvested when they show a withered state. On sunny days， leaves are cut off， and the roots are digged up， followed by shaking off the soil. After harvesting， the lateral roots of A. dahurica are removed， and the soil is washed off. After being dried for 1 to 2 d， the main roots are divided to the large， medium and small grades and exposed to blazing sun for a long time， respectively， when avoiding rain. The roots should be taken back and air-dried on rainy days， and then sun-dried on a sunny day. Otherwise， it will cause rot or black heart. The roots are turned over 1 to 2 times a day during the drying process. When the root tubers are beaten with a crisp sound， it indicates that they have been dried completely and are the finished product. The roots can also be dried in an oven， with the temperature controlled within 45 ℃. The oven-dried or sun-dried A. dahurica must have a moisture content of no more than 14%.

Packaging and storage

The processed products should be packaged in grades in time， and the packaging bags must be marked with the product name， specification （grade）， place of origin， batch number， packaging date， production unit， etc.， to ensure that the quality of A. dahurica meets the requirements of the Chinese Pharmacopoeia （2015 edition）. A ventilated， dry， dark， rodent-proof and moisture-proof sealed warehouse can be chosen for storage， and it should be checked regularly.

Conclusions

A. dahurica is a traditional Chinese medicinal material and a plant for both medicine and food purposes as announced by the National Health Commission. It has the effects of dispelling wind and relieving pain， expelling pus and dispersing swelling， clearing the nasal orifices， and eliminating dampness. It is mainly used to treat common cold due to wind-cold， headache， rhinitis， toothache， leukorrhea with reddish discharge， and sore swelling and pain， and can also used as a spice. In recent years， with the in-depth research on the chemical composition of A. dahurica， the medicinal value of A. dahurica has increased， the development of related products has increased， the market demand for A. dahurica has increased， and the cultivation of A. dahurica has also been vigorously developed， while the related standardized planting technology， production area processing and new product development research are backward， which restricts the development of A. dahurica industry. As an edible medicinal material， in addition to research on new drugs， its "Jian" brand and "food" brand products and daily chemicals should be developed more to expand its application market， increase the price of A. dahurica， and increase growers planting income; and it is necessary to carry out research on the standardized planting technology of A. dahurica， so that the planted products meet GAP certification， and the sustainable development of A. dahurica industry can be better promoted. This paper summarized the genetic relationship and production distribution of the different germplasm resources of A. dahurica. Combining the research of the research group on the cultivation technique of A. dahurica and referring to the research of other colleagues in the cultivation technique of A. dahurica， a systematic summary was made， hoping to provide reference and guidance for the research and industrial development of A. dahurica in Hunan area and promote the sustainable development of A. dahurica industry in Hunan.

References

[1] XU S， HU JQ， QUAN SC. The research and clinical application progress of the active components of Angelica dahurica[J]. China Pharmacy， 2005， 16（6）： 467-469. （in Chinese）

[2] XIA LM， YAO C. Research status of traditional Chinese medicine Angelica dahurica[J]. Research of Traditional Chinese Medicine， 2002， 18（5）： 56-58. （in Chinese）

[3] WANG ZS. Study on chemical composition and quality standard of Angelica dahurica[D]. Kaifeng： Henan University， 2008. （in Chinese）

[4] HUANG LQ， LYU DM， YANG B， et al. Development of research on germplasm resources of medicinal plants： Construction of core germplasm[J]. China Journal of Chinese Materia Medica，2005，30（20）：1565-1568. （in Chinese）

[5] ZHOU ZJ， WU XY， MENG YJ， et al. Research progress on medicinal plants breeding[J]. Lishizhen Medicine and Materia Medica Research， 2008， 19（7）： 1694-1698. （in Chinese）

[6] GUO DD. Investigation and evaluation of Angelica dahurica germplasm resources[D]. Chengdu： Chengdu University of Traditional Chinese Medicine， 2008. （in Chinese）

[7] GUO DD， MA YY， TANG L， et al. Genetic diversity of Radix Angelicae Dahuricae germplasmic resource based on ISSR analysis[J]. Chinese Traditional and Herbal Drugs2009，40（10）：1627-1630. （in Chinese）

[8] TANG SW， WANG MY， JIANG GH， et al. Study on genetic markers of Chuan Baizhi germplasm[J]. Modernization of Traditional Chinese Medicine and Materia Medica， 2004， 6（2）： 76-79. （in Chinese）

[9] MA YY， GUO DD， JIANG GH， et al. Investigation report on the germplasm resources of Angelica dahurica[J]. West China Journal of Pharmaceutical Sciences， 2009， 24（5）： 457-460. （in Chinese）

[10] WANG P. The research on the quality analysis and processing in production place of Angelica dahurica from Bozhou[D]. Hefei： Anhui University of Chinese Medicine， 2015. （in Chinese）

[11] WANG NH， QIN HZ， HUANG LQ， et al. Studies on original plant of traditional chinese drug "Bai Zhi” （Radix Angelicae Dahuricae） and its closely related wild plants II. karyological and pollen morphological studies on "Bai Zhi” and closely related wild plants[J]. China Journal of Chinese Materia Medica， 2001， 26（9）： 584. （in Chinese）

[12] HOU K. Evaluation of Sichuan Angelica dahurica resources and the influence of plant hormones on its growth and product quality[D]. Chengdu： Sichuan Agricultural University， 2013. （in Chinese）

[13] PAN ZH， QIN HZ， WU ZJ， et al. The karyotype research of Angelica dahurica and its classification[J]. Journal of Systematics and Evolution， 1985， 23（3）： 185. （in Chinese）

[14] HUANG LQ， WANG M， FU GF， et al. RAPD analysis of germplasm resources of Angelica dahurica[J]. China Journal of Chinese Materia Medica， 1999， 24（8）： 457-459. （in Chinese）

[15] YANG B， WANG M， CAO CY， et al. Molecular genetic study on the crude herb Baizhi and its source[J]. Chinese Pharmaceutical Journal， 2004， 39（9）： 654-657. （in Chinese）

[16] YUAN J， YIN PY， LI HL. Biochemical features and standardized cultivation technologv of Augelica dahurica[J]. Forest By-Product and Speciality in China， 2010（1）： 43-44. （in Chinese）

[17] XUE QF， ZHANG P， XU JL. Cultivation and pest control of Angelica dahurica[J]. Special Economic Animal and Plant， 2009（3）： 37-38. （in Chinese）

[18] PU SC， SHEN ML， TAN QS， et al. Technical regulations for standardized production of Nanchuan Angelica dahurica[J]. Modern Chinese Medicine， 2010， 12（9）： 13-17. （in Chinese）

[19] ZHAO DY， HAO QX， JIN Y， et al. Research advance on the biological characteristics and cultivation technologies of Angelica dahurica[J]. Modern Chinese Medicine， 2015， 17（11）： 1188-1192. （in Chinese）

[20] LIU XH. Angelica dahurica cultivation and management technology[J]. South China Agriculture， 2011， 5（9）： 30-31. （in Chinese）

[21] ZHAO ZM， ZHU YB. Artificial cultivation technology of medicinal plant Xingan Angelica dahurica[J]. Forest By-Product and Speciality in China， 2013（1）： 53-54. （in Chinese）