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Survey analysis of soil physicochemical factors that influence the distribution of Cordyceps in the Xiahe Region of Gansu Province

  • Shuling He , Lingfa Ma , Kentian Zhao EMAIL logo , Jingjun Yang and Yuwei Chang
Published/Copyright: April 19, 2017
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Open Life Sciences
From the journal Open Life Sciences Volume 12 Issue 1

Abstract:

In this paper, we studied the influence of soil physicochemical factors on the distribution of Cordyceps. We found that in the soil layers at different depths, the differences in pH values, total N (TN), total P (TP), available P (AP) and available K (AK) were not significant, but the differences in soil water content (WC), soil organic matter (OM), hydrolysable nitrogen (HN) and AK were significant. In the 5-10 cm layer, WC, pH and TP were significant factors that influence the distribution of Cordyceps - the number of Cordyceps was the largest and the characterization of the Cordyceps was also the best. TP was an important factor for the first principle component of the soil physicochemical properties that influenced the population distribution of Cordyceps. pH value was an important factor for the the second principal component of the soil physicochemical properties that influenced population distribution, and WC was an important factor for the third principal component, the soil physicochemical properties. This demonstrates that the requirement of Cordyceps for specific ranges of WC, soil acidity-alkalinity and AP in soil layers is very high.

Keywords: Cordyceps; soil environment; soil layer; physicochemical factor; habitat

1 Introduction

Cordyceps is a fungus form the Clavicipitaceae family. It comprises a stroma and sclerotia component resulting from the endoparasitic infection of swift moth larvae [1]. Dried fungi-larva sporocarp is highly valued in traditional Chinese medicine. Sources report that its ingestion has positive influence on human immune system. Cordyceps is primarily used for treating chronic cough, post-illness weakness, insomnia, emaciation, impotence, among other conditions [2-4]. The fungi life cycle is extremely sophisticated. It undergoes a relatively independent conidia stage as well as anascospore stage. This combined with host swift moth 5-6-year life cycle, 85% of which is underground, result in its extremely peculiar habitat requirements [5]. Increased market demand for Cordyceps makes its wild environment severely endangered [6]. Accordingly, recent research has focused on protecting the wild environment of this fungus.

Cordyceps has very strict requirements for ideal environmental conditions. It grows primarily on and around the Qinghai-Tibet Plateau, including Tibet, Qinghai, Sichuan, Gansu, Yunnan and other provinces. Normally it can be found on the alpine shrub meadows and alpine meadows at 3600-4500 m a.s.l. The suitable soil for its development is the fertile and loose alpine meadow soil with thick soil layer and moderate moisture. The occurrence of Cordyceps is characterized by a very clear zonal and vertical distribution pattern, which is consistent with its host distribution [7-10], which relies heavily on soil habitat. Rcent research has primarily focused on associated plants, altitude limits, eating patterns of swift moth larva and the relationship between soil microorganisms and the formation of Cordyceps [8-15]. So far, there are no reports on the soil microenvironment of Cordyceps within its distribution area in the Xiahe Region of Gansu Province. This paper presents new information about the relationship between Cordyceps population distribution and soil nutrients in this area. We hope the results will encouragethe environmentally-sensitive use of Cordyceps’ natural habitat and the sustainable use ofits semi-artificial cultivation.

2 Materials and methods

2.1 Research area

The research region included seven townships in Xiahe County of Gannan Tibetan Autonomous Prefecture: Ganjia Township, Damei Township, Zhayou Township, Tanggaang Township, Bola Township, Sangke Township, and Kecai Township. Xiahe County is located in the west edge of the southern part of Gansu Province. The county is surrounded by Zhuoni County in the east, Luqu County in the south, Linxia Prefecture in the north and Qinghai Province in the west. The altitude of this region is between 3000 m and 3800 m. The climate there is cold and humid with clear high plateau climate characteristics. The average temperature is 2.6℃, average rainfall is 516 mm, average frost-free period lasts 56 days and annual sunshine time is 2,296 h. The primary vegetation in the region includes Gentianaceae, Scrophulariaceae, Compositae, Rosaceae, Polygonaceae, and Gramineae, with a coverage rate of 78.5%. The soil thickness is between 32 and 42 cm and the soil types are primarily alpine meadow soil and dark felty soil.

2.2 Sample plot selection and soil sample collection

Under the guidance of local medicine farmers, forestry bureaus, Tibetan hospitals and Cordyceps markets, we chose sampling sites and established plots and visited them from early May to late June for four consecutive years 2013-2016. Plots were established in regions that reflected Cordyceps distribution characteristics and random sampling method was applied. Every year, we selected 10 new typical 10 m × 10 m plots (major plots). In each major plot, we selected 5 small 1 m × 1 m quadrats through the five-point method. 50 small sampling quadrats were created in this way. A deep hole (30-40 cm) was dug in teach quadrat. Soil samples at the layers of 0-5, 5-10, 10-15 and 15-20 cm were collected and 3 random samplings were conducted. The soil type was recorded. In the same sample site, soil samples in 5 quadrates at the layers of 0-5, 5-10, 10-15 and 15-20 cm were mixed, which was then used as the soil sample of corresponding quadrates. 20 soil samples were collected in this way. The samples were then sealed using the diagonal method. After being recorded of the type, they were carried to the laboratory in freshness protection packages for physicochemical property detection. We also recoded surrounding environment information, including soil type, altitude, slope gradient, grassland type and major vegetation.

2.3 Calculation of the amount of Cordyceps

We investigated and analyzed every quadrat, including the Cordyceps number, vegetation component, plant coverage, density and height, distribution status, growth status, soil type, and soil physicochemical properties. We selected 1 small quadrat from each major plot and dug the small quadrat to the depth of 50 cm. Then we inspected the number of host insects, weighed their biomass, measured the body length of the insect and the length of stroma and calculated its resource mass. For some major plots, due the severe damages caused medicine farmers digging, the number of Cordyceps could not be accurately calculated. In such cases, we could only estimate it using the number of holes left after digging.

2.4 Analysis of the soil physiochemical properties

(1) The soil organic matter (OM) was tested using the electric sand bath and potassium dichromate titrimetric method. (2) The total nitrogen (TN) was tested using the Semimicro Macro Kjeldahl method. (3) The total phosphorus in soil (TP) was tested using the NaOH melting method —Mo-Sb anticolorimetric method. (4) The total potassium (TK) was tested using the NaOH melting method- flame photometer method. (5) The hydrolysable nitrogen (HN) was tested using the alkaline hydrolysis diffusion method. (6) The rapidly-available phosphorus (AP) was tested using the sodium bicarbonate method. (7) The rapidly-available potassium (AT) was tested using the ammonium acetate—flame photometer method (1 mol/L neutral NH4OAc extraction). (8) The water content (WC) was tested using the oven drying method. (9) The soil pH value was measured using the potentiometry method.

2.5 Data analysis

We used DPS 7.5 to analyze, process and conduct principle component analysis of the data. After we obtained the principle component, we conducted a hierarchical cluster analysis on it to confirm the principle component of the soil nutrient selection of the Cordyceps. We then analyzed the influence of water content, pH value, organic matter, total nitrogen, hydrolysable nitrogen, total phosphorus, AP and AT on the population distribution of Cordyceps.

3 Results

3.1 Influence of soil depth on the characteristics and number of Cordyceps

We found that the numbers of larva differed greatly in different soil depths. From Table 1, we can see that as the soil depth increased, the number of Cordyceps first increased and then decreased and so did the diameter and size of Cordyceps. The number of Cordyceps was the largest in the soil layer 5-10 cm and was the smallest in soil layer 20-25 cm. In addition, the number of Cordyceps in soil layer 20-25 cm was significantly 85.67% less than that in the soil layer 5-10 cm. This indicates that Cordyceps primarily lives in soil layer 5-10 cm in the Xiahe Region in Gannan Tibetan Autonomous Prefecture and that the soil layer over 20 cm is not suitable for Cordyceps to grow, as suggest the sources [10-13].

Table 1

Influence of soil depth on the characteristics and number of Cordyceps.

Soil layer [cm]Larva length [cm]Larva diameter [mm]Stroma length [cm]Stroma diameter [mm]Larva number
0-52.31 ± 0.12bB3.67 ± 0.12aA2.55 ± 0.09bB2.46 ± 0.47bB2.31 ± 0.12bB
5-102.67 ± 0.13bB4.35 ± 0.17aA3.27 ± 0.43bB2.78 ± 0.44bB3.84 ± 0.38cB
15-203.84 ± 0.38cB4.97 ± 0.34bB4.53 ± 0.23bcB6.33 ± 0.44aA2.67 ± 0.13bB
20-250.55 ± 0.12abAB0.65 ± 0.13aA0.46 ± 0.13bcAB0.36 ± 0.09cB0.55 ± 0.12abAB

3.2 Environmental characteristics of Cordyceps main distribution areas

Table 2 suggests that in Zhayou Township,Tanggaang Township, Bola Township, Sangke Township and Kecai Township and Nawu Township which are located at an upper slope, over 3000 m in altitude and having an annual precipitation of 340 mm, the number of Cordyceps is more,In contrast, the number of Cordyceps in Ganjia Township and Damei Township was less. Vegetations in the habitat primarily include Gentianaceae, Scrophulariaceae, Compositae, Equisetaceae, Rosaceae, Polygonaceae and Gramineae. Soil types primarily include wet and dark felty soils, dark felty soils, felty soils, thin felty soils, cold and brown calcic soils, cold calcic soils, and cold and dark calcic soil. The primary plants were Kobresia myosuroides, Potentilla anserine, Equisetum arvense, Bistorta macrophylla, Polygonum viviparum, Potentilla fruticosa. The vegetation coverage rate was 75% and the soil thickness was 30-40 cm.

Table 2

Environmental characteristics of the main distribution areas of Cordyceps.

Environmental characteristicsGanjia TownshipDamei TownshipZhayou TownshipTanggaang TownshipBola TownshipSangke TownshipKecai Township
Altitude2950278030203210301231003000
[m a.s.l.]
Annual average415432332510630450340
precipitation
[mm]
Annual average3.44.60.32.81.91.21.6
temperature
[℃]
Cordyceps number1-20-22-52-62-51-52-5
[item/m2]
Soil altitudinalDark brown soilDark brown soilBrown feltyDark brown soilBrown soilBrown soilBrown soil
zonesoil
Vegetation typeGentianaceaeCompositaePapaveraceaeGramineaePrimulaceaeGramineaeGentianaceae
ScrophulariaceaePapaveraceaeGramineaeRosaceaeGramineaeCompositaeGramineae
CompositaeLeguminosaeRosaceaeEquisetaceaeRosaceaeRosaceaeRosaceae
EquisetaceaePolygonaceeCompositaeCompositaePolygonaceaePolygonaceaeCompositae
RosaceaePolygonacae
Soil typeWet and dark feltyDark feltyDark feltyThin feltycold and brownCold calcicCold and dark
calciccalcic
Slope positionUpperUpperUpperUpperMiddleMiddleUpper

3.3 Physicochemical properties of soils at different layers

We conducted single-factor variance analysis on the physicochemical properties of the soil from 50 quadrates, the results suggests (Table 3) that the differences in pH value, TN, TP, AP and AK of the three soil layers were not significant (P > 0.05) but that the differences in WC, OM, HN and TK were extremely significant (P < 0.01). The content of OM, HN and AK in the soil in the major distribution area of Cordyceps was high. The pH value gradually decreased as the soil layer depth increased. Other soil nutrient indicators also exhibited this inverse relationship.

Table 3

Investigation on the soil physicochemical properties of different soil layers.

VariablesSoil layer [cm]
0-55-1010-1515-20
Water content (WC)35.08 ± 0.07cC33.76 ± 0.06bB32.32 ± 0.04aA28.14 ± 0.05dD
Organic matter (OM)40.01 ± 0.05bB45.28 ± 0.08aA36.32 ± 0.07cC30.35 ± 0.10dD
pH6.85.00 ± 0.08aA6.88 ± 0.12aA6.89 ± 0.05aA6.90 ± 0.07aA
Total nitrogen (TN)4.21 ± 0.05aA4.29 ± 0.07aA4.18 ± 0.028aA4.16 ± 0.11aA
Hydrolysable nitrogen (HN)38.72 ± 0.05bB42.05 ± 0.08aA35.21 ± 0.07cC32.20 ± 0.09dD
Total phosphorus (TP)1.51 ± 0.07aA1.53 ± 0.06aA1.52 ± 0.06aA1.48 ± 0.09aA
Avidly-available phosphorus (AP)18.39 ± 0.05aA18.47 ± 0.05aA18.34 ± 0.05aA18.34 ± 0.06aA
Total potassium (TK)4.70 ± 0.07bB4.89 ± 0.05aA4.62 ± 0.06cC4.47 ± 0.06dD
Available potassium (AK)142.01 ± 0.06aA143.65 ± 0.10aA142.79 ± 0.05aA144.82 ± 0.09aA

Note: Lowercase letters indicate significance at P<0.05 Uppercase letters indicatesignificance at P<0.01.

3.4 The eigen values and variance contribution rates of relevant coefficients in the principal component analysis of soil physicochemical properties

From the principal component analysis (Table 4), we can see that that the eigen values of the first two principle components were K1 = 6.4908 > 1.0000 and K2 = 1.5868 > 1.0000 and the contribution rate was the highest; the accumulative contribution rate was 200% > 85%, indicating that the first three principal components basically included 100% of the information of the nine parameters (WC, OM, pH, TN, HN, TP, AP, TK and AK) in soil layers 0-5, 5-10, 10-15 and 15-20 cm.

Table 4

The eigen values and variance contribution rates of soil physicochemical properties.

Principal componentEigenvalueContribution rate /%Accumulative contribution rate /%
16.49172.12072.120
21.58717.63289.752
30.92210.248100

3.5 Factor rotation of soil physicochemical properties

On the basis of principal component analysis, we selected the first three principal components from the factor matrix after varimax orthogonal rotation and used them to calculate the eigenvector of variable. Table 5 suggested that AP had the highest correlation coefficient in the first principal component, indicating that an important factor influencing the soil physiocochemical properties was AP [7-10]. In the second principal component, pH had a relatively high correlation coefficient, which reflected the strict requirement of Cordyceps on the acidity-alkalinity. In the third principal component, WC had the relatively high correlation coefficient, suggesting that WC was an important factor that influenced the distribution of Cordyceps in soil.

Table 5

Transposed matrix of the variable eigenvector of soil habitat of Cordyceps.

VariableThe first eigenvectorThe second eigenvectorThe third eigenvector
Water content (WC)0.2567-0.19850.9459
Organic matter (OM)0.8609-0.30440.4077
pH-0.31130.9501-0.0197
Total nitrogen (TN)0.9743-0.04560.2207
Hydrolysable nitrogen (HN)0.8942-0.33030.3023
Total phosphorus (TP)0.5890-0.16650.7908
Avidly-available phosphorus (AP)0.9951-0.06950.0709
Total potassium (TK)0.8972-0.19920.3941
Available potassium (AK)-0.00350.8505-0.5259

3.6 The influence of soil physicochemical properties on the distribution of Cordyceps

After standardizing the original data using the clustering analysis method, we conducted a statistical analysis of the influence of the physicochemical properties factors of the three layers of soil on the distribution of Cordyceps. The results showed that in soil layer 0-5 cm, AP and TK had an extremely significant influence on the distribution of Cordyceps, pH and TN had a significant influence and the first four factors that influence the distribution of Cordyceps in this layer were respectively AP > TK > pH > TN. In soil layer 5-10 cm, PH and AP had an extremely significant influence on the distribution of Cordyceps, WC and OM had a significant influence and the first four factors that influence the distribution of Cordyceps in this layer were respectively AP > pH > WC > TK. In soil layers 15-20 cm and 20-25 cm also the first four factors that influence the distribution of Cordyceps in these layers were AP > pH > WC > TK (Table 6).

Table 6

Influence of the soil physicochemical properties on the distribution of Cordyceps.

VariableSoil layer/cm
0-55-1015-2020-25
Water content (WC)0.68bB0.79bA0.67cC0.62cC
Organic matter (OM)0.87aA0.89aA0.81bA0.79bA
pH0.72bA0.74bB0.74bB0.76bA
Total nitrogen (TN)0.74bA0.89aA0.72bB0.71bB
Hydrolysable nitrogen (HN)0.95aA0.93aA0.92aA0.90aA
Total phosphorus (TP)0.92aA0.95aA0.91aA0.88aA
Avidly-available phosphorus (AP)0.95aA0.99aA0.95aA0.95aA
Total potassium (TK)0.63bB0.72bB0.82bA0.82bA
Available potassium (AK)0.72aA0.97aA0.95aA0.93aA

4 Discussion

The research results suggested that the water content in soil layer 0-5 cm was the highest, which was likely associated with the precipitation in that region. In contrast with Maqu and Luqu, the Xiahe Region was rather dry. The water content in soil layer 15-20 cm was the lowest. The larva of swift moth preferred high humidity (soil humidity 40%-46%) [6]. Through the analysis of the physicochemical properties of the soil, we found that the soil in which Cordyceps were growing in were primarily wet and dark felty soil and wet felty soil, whose humidity is compatible with biological needs. Other physicochemical properties in the soil such as pH value, total N (TN), total P (TP), available P (AP) and available K (AK) in different layers showed no significant differences. In this study, we found that the differences in WC, OM, HN and TK in the three soil layers were extremely significant. The survey data about the environmental characteristics of the distributed area suggested that the Cordyceps were primarily distributed in the upper slope or middle slopes of alpine shrub meadow and alpine meadow soil layers at an altitude above 3,100 meters.

This study provides supporting evidence that winter larvae primarily live in the soil layer 5-10 cm. Since the Cordyceps host primarily spends their life cycle in the soil, its habitat directly influences the population distribution of Cordyceps. Through this study, we can see that in the Xiahe Region of Gansu Province, AP, pH and WC are the major soil factors that influence the growing and distribution of Cordyceps. The host larva growth may also be subject to the influence of other biological factors or vegetation types which requires further study.

As the soil depth increased, the number, size and diameter of Cordyceps first increased and then decreased. The number of Cordyceps was the largest in soil layer 5-10 cm and the characterization was the best. In this region, the soil was slightly alkaline; and the differences in pH values in the three soil layers were not significant, which was consistent with the research result by Wu et al. [5].

5 Conclusion

Based on three years continuous tracking, on-site visit, field survey and statistical analysis in the Cordyceps distribution area of the Xiahe Region, we have found that the number of Cordyceps in soil layer 5-10 cm was the largest among the four soil layers in our survey and that the characterization of the Cordyceps was also the best in this layer. We also found that the physicochemical parameters had a great influence on the population distribution and survival of Cordyceps. Through principle component analysis and clustering analysis, we concluded that the rank of the parameters’ influence on the distribution of Cordyceps was AP > pH > WC > TK > TN > OM > HN > TP > AK. We hope that theses results will help in sustainable harvesting practices by medicinal farmers in order toprotect Cordyceps habitat and its semi-artificial cultivation. We also hope that the results will help in sustainable use of wild resources and provide theoretical references for protecting the grasslands and maintain ecological balance in the Gannan Plateau.

Funding

Funding was provided by Science and technology support of Gansu Province (1604NKCA073) and Project of Natural Science Foundation of Gansu Province (1506RJZP001).

  1. Conflict of Interest: Authors state no conflict of interest.

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Received: 2016-2-16
Accepted: 2016-9-13
Published Online: 2017-4-19

© 2017 Shuling He et al.

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

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https://doi.org/10.1515/biol-2017-0008
Keywords for this article
Cordyceps; soil environment; soil layer; physicochemical factor; habitat
Creative Commons
BY-NC-ND 3.0

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