Investigation on the Resources of Paris Polyphylla var. yunnanensis

Shicai WU, Guo LI, Qiping WANG, Wenli YANG, Xuemei CHEN, Jian FENG, Ganghua LI, Shaoshan ZHANG

1. Xingwen People’s Hospital, Yibin 644000, China; 2. Xingwen Hospital of Chinese Medicine, Yibin 644000, China; 3. Institute of Qinghai-Tibetan Plateau, Southwest Minzu University, Chengdu 610041, China

Abstract In this study, the resource distribution, morphological characteristics, artificial cultivation and market circulation of Paris Polyphylla var. yunnanensis and its polygerm varieties in Yunnan were investigated by referring to the research literature, field investigation and our planting experience. It is found that P. polyphylla var. yunnanensis is widely distributed in Yunnan Province, realizing large-scale artificial cultivation, while the wild resources of polygerm varieties are almost on the verge of extinction and are in urgent need of protection.

Key words Paris Polyphylla var. yunnanensis, Resource distribution, Cultivation, Investigation

1 Introduction

Parissp. is a plant belonging to Liliaceae family, and its medicinal part is underground rhizome[1]. It is the core raw material of many kinds of proprietary Chinese medicines, such as "Gongxuening capsule", and "Yunnan Baiyao aerosol". Since SARS in 2003, the consumption of Rhizoma Paridis has soared. At present, the consumption of Rhizoma Paridis far exceeds the total growth of all wildParisspp. in China, so that it can not meet the market demand for Rhizoma Paridis medicinal materials[1-5].

According to the morphological characteristics and evolution trend of the genusParis, Li Hengetal. put forward the classification system ofParisspp., and divided it into 2 subgenera, 8 groups, 27 species and more than 10 variants. There are 30 species and variants in China[2,6], and they are mainly distributed in southwest China and South Asian countries. The previous edition ofPharmacopoeiaofthePeople’sRepublicofChinaonly containsParispolyphyllaSmith var. Yunnanensis (Franch.) Hand.-Mazz. orParispolyphyllaSmith var. chinensis (Franch.) Hara[7]. PolygermP.polyphyllavar. Yunnanensis (polygerm varieties) is a new strain with typical botanical characteristics ofParisPolyphyllavar. yunnanensis, which has been discovered in recent years. For example, the upper part of the petal is often enlarged into a narrow spoon shape with a width of 2 mm, the protruding part of the connective is obvious, with a length of 1-2 mm[1]. However, compared withP.polyphyllavar. yunnanensis, it has a large number of cluster buds, each of which can be used as a cultivation material. Therefore, it can propagate rapidly and increase production significantly. Gaoligong Mountains area is located in the west of the southern section of National Nature Reserve Gaoligong Mountains, and the hybridization with external species is naturally cut off[8]. Therefore,P.polyphyllavar. yunnanensis (polygerm varieties) in this area is quite different fromP.polyphyllavar. yunnanensis (polygerm varieties) in other areas in terms of plant morphological characteristics, shape and number of cluster buds and so on.P.polyphyllavar. yunnanensis (polygerm varieties) has been paid more and more attention to by researchers, especially the cultivation enterprises and growers because of its many advantages.

At present, the resources ofParisspp. in southwest China have been investigated, and a lot of studies have been done on the cultivation ofParisspp., especiallyP.polyphyllavar. yunnanensis andP.polyphyllavar.chinensis. The research results provide a certain basis for the evaluation ofParisresources and the development of the industry[8]. This study focuses on investigating the latest resources distribution ofP.polyphyllavar. yunnanensis and its polygerm varieties and summarizing the cultivation techniques in different regions, in order to provide scientific basis for variety protection and industrialization development ofP.polyphyllavar. yunnanensis and its polygerm varieties.

2 Materials and methods

2.1 MaterialsDigital camera: used to take pictures of plant morphology, habitat, specimen,etc. The GPS tool: Magellan Triton, used to record the geographic information of the collection area. Digging tools: hoes, shovels,etc., used to collect samples of Rhizoma Paridis. Specimen clip/paper: used for specimen pressing. Collection recording paper, needle and thread, specimen tag,etc.: used to record specimen information. Experimental notes,etc.: used to record investigation information.

2.2 Methods

2.2.1Query of literature, books and other materials. Through CNKI and Wanfang Database, we searched the relevant data and literature on the resource distribution, quality evaluation, ecological environment distribution and cultivation techniques ofP.polyphyllavar. yunnanensis and its polygerm varieties. By consulting the specimens collected by the herbarium with richParisspp. in Southwest China (YUKU, KUN, CDBI, CDCM, WCU, SM, SZ,etc.), we had a preliminary understanding of the resource distribution ofP.polyphyllavar. yunnanensis and its polygerm varieties, and then formulated a detailed investigation plan.

2.2.2On-the-spot investigation. The period from April to July is the flowering period ofP.polyphyllavar. yunnanensis and its polygerm varieties. During this period, we investigated the field and planting bases in the main producing areas, collected specimens and samples, visited local growers, veteran practioners of TCM, and consulted relevant materials and literature in the later period, to find out the local resource distribution, reserves, artificial cultivation techniques and local introduction and cultivation ofP.polyphyllavar. yunnanensis and its polygerm varieties.

Yunnan area: it has been verified that the resources ofP.polyphyllavar. yunnanensis and its polygerm varieties are mainly distributed in the natural basin of Nujiang River, Dulong River and Gaoligong Mountains. In terms of administrative division, it is in Kunming City, Yunnan Province-Chuxiong Autonomous Prefecture-Dali Bai Autonomous Prefecture-Lijiang City-Diqing Tibetan Autonomous Prefecture-Nujiang Prefecture-Baoshan City and other western Yunnan regions. Therefore, the selected areas in this survey are Luquan County of Kunming City, Wuding County of Chuxiong Autonomous Prefecture, Yuanmou County of Chuxiong Autonomous Prefecture, Gongshan County of Nujiang Prefecture, Diqing Tibetan Autonomous Prefecture, Huaping County of Lijiang City, Ninglang County of Lijiang City, Yongsheng County of Lijiang City, Weixi County of Lijiang City, Weishan County of Dali Bai Autonomous Prefecture, Longling County of Baoshan City, Changning County of Baoshan City, and Tengchong City. The area is connected with the Qinghai-Tibet Plateau in the north and China-Myanmar in the south, showing a north-south direction and spanning five latitudinal zones. It is the only area on earth that has preserved a large area of zone of transition from humid tropical forests to temperate forests so far, and it is an area with extremely rich and precious biodiversity in the world. Tengchong City is located in the upper part of the southern section of Gaoligong Mountains, and is closely surrounded by Gaoligong Mountains, and the species is almost isolated from the outside world. The characteristics of plant morphology and the number and shape of cluster buds inP.polyphyllavar. yunnanensis (polygerm varieties) in this area are very different from those in other areas, so the resource status ofP.polyphyllavar. yunnanensis (polygerm varieties) in this area is the focus of the investigation. Lijiang City, Dali Prefecture and Tengchong City are the early cultivation areas ofP.polyphyllavar. yunnanensis and its polygerm varieties, so the three places are the key areas of cultivation technology investigation.

Sichuan area: Longmen Mountain-Qionglai Mountain-Jiajinshan Mountain-Erlang Mountain-Emei Mountain-Daliang Mountain. From north to south, the investigation areas are Qingchuan County, An County, Beichuan County, Wenchuan County, Pengzhou City, Chongzhou County, Dayi County, Lushan County, Tianquan County, Luding County, Kangding County, Hongya County, Pixian County, Emeishan City, Leshan City, Pingshan County and Panzhihua City. Geologically, this area is the transitional area from Hengduan Mountains to Sichuan Basin, with great differences in climate and rich types of vegetation, which is one of the traditional distribution centers ofParisspp..

Guizhou area: Guiding County, Huishui County and Hezhang County, Qiannan Autonomous Prefecture, Guizhou Province. The area has a large elevation gap, mild climate and fertile soil, and is also one of the traditional distribution centers ofParisspp.

3 Results and analysis

3.1 Original plant identificationWhen Takhtajan studied the groupP.polyphyllain 1983, because most of the species indexes used were variable characters, such as sepal shape, stamen number, petal shape and leaf base shape, and all the variants of this species were promoted to species. For example,P.polyphyllaSmith var.Yunnanensiswas upgraded toP.polyphyllaSmith Daiswa.Yunnanensis, andP.polyphyllaSmith var. chinensis was promoted toP.polyphyllaSmith D. chinensis[2]. After consulting the specimens collected from various places in Yunnan, Sichuan and Guizhou, as well as collecting and observing plants on the spot, in accordance with the research results of Takhtajan, researcher Li Heng classifiedP.polyphyllavar. yunnanensis as a variety ofP.polyphylla, which was published inParisGenusPlantin 1998, and formally identified it asP.polyphyllaSmith var.yunnanensis(Franch.) Hand.-Mazz.[2]. The focus of this study-P.polyphyllavar. yunnanensis (polygerm varieties) was approved by Sichuan Provincial Crop Variety Approval Committee as the new material "YDCL03" on August 28, 2014. Its basic source isP.polyphyllaSmith var. yunnanensis (Franch.) Hand.-Mazz. Therefore, this study only adds a suffix (polygerm varieties) to PolygermP.polyphyllavar. yunnanensis (Franch.) Hand.-Mazz. for the purpose of making a distinction.

In the course of investigation, specimens ofP.polyphyllavar. yunnanensis and its polygerm varieties were collected and pressed, and their geographic information was recorded. In accordance with literature, photos and measurement data, this paper complemented the morphological description ofP.polyphyllavar. yunnanensis and its polygerm varieties respectively. The morphological characteristics ofP.polyphyllavar. yunnanensis are as follows:

The rhizome is stout, 11 cm long and 1.0-4.2 cm thick; the stem is 17-84 cm high, glabrous. Leaves (5-12) are green, thick, obovate-oblong, obovate-elliptic, ovate-lanceolate. The base is wedge-shaped to rounded, 3-10 cm long and 1.2-4.5 cm wide, often with a pair of distinct basal veins, and the petiole is 0-2 cm long. The pedicel is 1.8-3.5 cm long, the fruit stage is obviously elongated, and the number of flower base is 3-7. Sepals are green, lanceolate, 2.2-8 cm long, sometimes shortly clawed, 0.1-0.9 cm wide, yellowish green or green, sometimes stem yellowish green, upper purple. Petals are green or yellowish green, filiform or ribbon-shaped, longer than or as long as sepals, often enlarged distally into a narrow spoon 0.2-0.5 cm wide. Stamens have 2-4 whorls (occasionally deviation of 1), 0.9-1.8 cm long, and filament is 0.2-0.7 cm long. The anther is 0.5-1.2 cm long, the protruding part of the connective is obvious, 0.1-0.3 cm long, or slightly dilated. Ovary is purple, smooth or tuberculate, angled or winged, 1-loculed. Placenta (3-7) is flat or raised toward the ventricle. Stylopodium is purple, thickened, often angled and discoid. Style is purple, 0-0.3 cm long. Stigma is purple, 0.4-1.1 cm long, erect when flowering, revolute in fruiting. Fruit is subglobose, yellowish green or green, irregularly dehiscent at maturity, 1.5-4.4 cm in diameter. Seeds are ovoid or reniform, with bright red or reddish exotesta. The flowering period is from April to July and the fruiting period is from September to November.

The morphological characteristics ofP.polyphyllavar. yunnanensis (polygerm varieties)-"new material YDCL03" are as follows:

Plant is 15-73 cm high, rhizome is cylindrical, with many digitate rhizomes. Leaves (6-12) are ovate-lanceolate, petiole is 0.5-2.0 cm long, and calyx (3-6) is lanceolate or ovate-lanceolate, 3-8 cm long. Petals (3-6) are strip-shaped and erect, with a width of up to 6.5 mm above the middle and a length half of or nearly equal to the length of the calyx: the stamens (6-12), the anthers are 1.0-1.5 cm long, the filaments are very short, and the protruding part of the connective is about 0.5-2.0 mm; the ovary is globose, the style is thick and short, the florescence is from June to July, and the fruiting period is from September to October.

3.2 Investigation of plant resources ofP.polyphyllavar. yunnanensis and its polygerm varieties

3.2.1Yunnan area.P.polyphyllavar. yunnanensis is distributed almost all over Yunnan Province. It mainly grows in natural basin of Nujiang River, Dulong River, Gaoligong Mountains, Yulong County and Ninglang County of Lijiang City in western Yunnan. It is also cultivated in Qujing City, Zhaotong City and other places in central and eastern Yunnan. In this survey,P.polyphyllavar. yunnanensis (polygerm varieties) is mainly distributed in Lijiang City, Yunnan Province (such as Yunnan Baiyao "Yunquan No.1" planting base). It is cultivated in small quantities in Jinghong City, Xishuangbanna Dai Autonomous Prefecture. It is cultivated in small quantities in Yunlong County and Weishan County of Dali Bai Autonomous Prefecture (such as Jiahe wild R. Paridis breeding and planting base), and Liuku Town, Lushui County, Nujiang Prefecture (most of the provenances in this area come from Tengchong City, a few from Lijiang City and Dali Prefecture). Yunlu Biotechnology Co., Ltd. and Funong Traditional Chinese Medicine Professional Cooperative, Longling County, Baoshan City, have large-scale cultivation (provenance comes from Dali Prefecture and Tengchong City), and Mingguang Town and Jietou Town in Tengchong City also have large-scale cultivation. Among the above-mentioned areas, because of its special geographical location, the biological characteristics ofP.polyphyllavar. yunnanensis and its polygerm varieties in Tengchong are different from those of the typical R. Paridis described in the literature, and different from those of the banded and wide-petalP.polyphyllavar. yunnanensis which is widely distributed in other areas. Most of the characters are betweenP.polyphyllavar. yunnanensis andP.forrestii(Takht) H. Li. (such as slender petals), and betweenP.polyphyllavar. yunnanensis andP.daliensis(such as bulging part of the connective).

3.2.2Guizhou area. There is typical Rhizoma Paridis Yunnan-ensis in Anshun, Huishui, Guiding and other areas of Guizhou, while the morphological characteristics ofP.polyphyllavar. yunnanensis in Hezhang County of Guizhou Plateau are similar to those ofP.polyphyllavar. chinensis which is widely distributed in this area. In this investigation,P.polyphyllavar. yunnanensis (polygerm varieties) was not found to be distributed in Guizhou.

3.2.3Sichuan area.P.polyphyllavar. yunnanensis is mainly distributed in western Sichuan and Panxi region.P.polyphyllavar. yunnanensis was collected by the research group in Xichang City, Yanyuan County, Wolong Town (Wenchuan County), Muli Tibetan Autonomous County and other places. Among them, the morphological characteristics ofP.polyphyllavar. yunnanensis in Yanyuan County and Muli County have many similarities with the widely distributedP.pubescensin this area, such as leaf veins and petioles densely covered with white short hairs. There is no large-scale distribution ofP.polyphyllavar. yunnanensis (polygerm varieties) in Sichuan. It is only introduced and cultivated in Pengzhou City, Pixian County of Chengdu City and Panzhihua City, and the provenances come from Tengchong City, Yunnan Province.

The mountain area of Gaoligong Mountains has unique geographical environment and climatic conditions, which is extremely rich in biodiversity in the world. Tengchong is located to the west of the southern section of Gaoligong Mountains, separated by the Nujiang River Valley to the east, Longling County to the south, Lushui County to the north, and Myanmar to the west. The temperature is suitable for the whole year, and as the hot and humid southwest monsoon is blocked by Gaoligong Mountains, the precipitation is richer than that in the neighboring areas. There are more than 2000 kinds of higher plants in the area, known as "natural plant source" and "plant gene bank".P.polyphyllavar. yunnanensis (polygerm varieties) in Gaoligong Mountains area represented by this city is obviously different from that in other places, and the comparison results are shown in Table 1.

4 Discussion

4.1 The reserves and development trend of wildP.polyphyllavar. yunnanensis and its polygerm varietiesDue to the increasing demand for Rhizoma Paridis in the pharmaceutical industry, wildParisspp. resources are increasingly scarce. The reproduction ofParisspp. is threatened to varying degrees, and the current living condition is worrying. The distribution ofP.polyphyllavar. yunnanensis and its polygerm varieties were hardly found in the field investigation in Yunnan, Sichuan and Guizhou. If it is not strictly protected, it can be predicted that this variety will be on the verge of extinction.

Table 1 Comparison of Paris Polyphylla var. yunnanensis (polygerm varieties) between Tengchong City and other areas

4.2 The basic situation of artificial cultivation ofP.polyphyllavar. yunnanensis and its polygerm varietiesAccording to the statistics of Yunnan Province, the cultivation area ofParisspp. in Yunnan Province is about 3 333.33 ha, and it is dominated byR.ParidisYunnanensisexcept for a small number of varieties such asP.forrestii(Takht) H. Li. andP.pubescens. The cultivation area ofP.polyphyllavar. yunnanensis (polygerm varieties) unique to Tengchong area is about 33.33 ha, which is mainly distributed in Mingguang Town, Jietou Town and Diantan Town of Tengchong City. In Sichuan area, except for cultivation in small quantities in Yanjin County near Yunnan, the rest of the area is limited to experimental introduction. Guizhou is also the traditional distribution place ofP.polyphyllavar. yunnanensis, but the artificial cultivation in this area started late with poor foundation. At present, large-scale artificial planting has not been formed.

4.3 The basic situation of market demand and trade ofP.polyphyllavar. yunnanensis and its polygerm varietiesAccording to the statistics of Yunnan Rhizoma Paridis Planting and Development Co., Ltd., the demand of national pharmaceutical companies for Rhizoma Paridis is about 15 000 t, and the demand for prescription of traditional Chinese medicine is 1 000 t. If so, the annual consumption of roots and stems ofParisspp. in the whole country should be 16 000 t. On the basis of 3 750 kg of Rhizoma Paridis medicinal material per hectare, 4 266.67 ha of Rhizoma Paridis products are needed every year. In addition, it takes more than 6 years for seed propagation to obtain Rhizoma Paridis medicinal materials, so the planting land should be increased by 6 times to ensure the normal supply of Rhizoma Paridis, that is, a demand for 25 600 haParisspp. in total. After nearly 10 years of operation, the planting area ofParisspp. in 2014 was only 133.33 ha. In order to meet the demand of the current traditional Chinese medicine market, it will take 19 years if 133.33 haParisspp. can be expanded every year, even if it is supported by the government and all sectors of the community.