Arbuscular mycorrhizal colonization of a Cd hyperaccumulator Viola baoshanensis at Baoshan Pb/Zn Mine

doi:10.21203/rs.3.rs-2008733/v1

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Arbuscular mycorrhizal colonization of a Cd hyperaccumulator Viola baoshanensis at Baoshan Pb/Zn Mine

https://doi.org/10.21203/rs.3.rs-2008733/v1

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Despite great potential for arbuscular mycorrhizal (AM) fungi associated with heavy metals (HMs) hyperaccumulators in the phytoremediation of contaminated sites, rather limited information is available in literature about the community structure of AM fungi (AMF) in the rhizosphere of hyperaccumulators in nature. A field survey was conducted to investigate the colonization status and community structure of AMF of Viola baoshanensis, a Cd hyperaccumulator, growing at Baoshan Pb/Zn Mine. Shoot/root ratios of 1.78 for Cd, and 2.57 for Zn in V. baoshanensis indicate that these two metals were preferentially transported from roots to shoots, whereas Pb was mainly stored in roots. The roots of V. basshanensis were extensively colonized by AMF with M% of 69.1%, and A% of 46.9%, whereas mycorrhizal colonization was not affected by concentrations of Cd, Zn and Pb in the soil. 15 AMF species ( 5 Glomus, 3Rhizophagus,2 Claroideoglomus, 2 Septoglomus, and each one of Ambispora, Funneliformis, and Sclerocystis ) were identified by a morphological method. The dominant AMF genus was Glomus, showing high tolerance to excess Cd, Zn and Pb, while Glomus ambisporum and Claroideoglomus etunicatum were the most abundant species in the rhizosphere of V. baoshanensis.

Mycology

Arbuscular mycorrhizal fungi

Hyperaccumulator

Cadmium

Lead

Zinc

Many mining sites were characterized by great toxicity of HMs, and poor nutrients and soil structure leading to destroyed vegetation cover ( Wong 2003; Orłowska et al. 2005 ), meanwhile revegetation of these sites is urgent in order to inhibit HMs contamination of the surrounding soil via erosion and runoff (Shetty et al. 1994; Vieira et al. 2018). However, there are many plants called metallophytes endemic to the contaminated soils have developed tolerance mechanisms to HMs (Perrier et al. 2006; Antosiewicz et al. 2008). The use of these plants to remediate the HMs contaminated sites are known as phytoremediation, which is environmentally friendly and cost-effective compared with conventional methods (McGrath &Zhao 2003; Arthur et al. 2005; Rajkumar et al. 2012).

Hyperaccumulators, a particular group of metallophytes, are able to accumulate HMs in shoots at extraordinary high levels and can be used in one of the key processes of phytoremediation called phytoextraction (Reeves & Baker 2000; Yang et al. 2019). An ideal candidate for efficient phytoextraction requires rapid growth and big shoot biomass of the plant (Wei et al. 2014 ). Nevertheless, hyperaccumulators often have slow growth and low biomass ( Khan et al. 2000), optimization of their fitness and growth need to consider the symbiotic microorganisms in soil ( Sánchez-Castro et al. 2017; Wang et al. 2021) .

The most important symbiotic partner is AMF, as they can form symbiosis with most of higher plants, providing a direct connection between soil and plant roots (Smith & Read 1997). AMF can efficiently improve water and minerals uptake of plants, therefore plants are widely colonized by AMF in many polluted soils (Shetty et al. 1994; Pawlowska et al. 1996; Del Val et al. 1999; Whitfield et al. 2004). However, until Berkheya coddii was found to be colonized by AMF (Turnau & Mesjasz-Przybylowicz 2003 ), hyperaccumulators had been thought as non-mycorrhizal (Leyval et al. 1997), since there may be a trade-off between hyperaccumulation and mycorrhization because of both requiring carbon investment from plants (Audet 2013). Thereafter, several mycorrhizal hyperaccumulators have been documented (Regvar et al. 2003; Vogel-Mikuš et al. 2005; Perrier et al. 2006; Wu et al. 2007; Rashid et al. 2009; Wei et al. 2014 ). It has been demonstrated that AMF can increase biomass and survival of hyperaccumulators, thus showing a great potential for phytoextraction (Al Agely et al. 2005; Orłowska et al. 2013 ).

However, there is a lack of clear elucidation about AMF role in hyperaccumulation of HMs, as the effects of AMF on metal uptake are diversified (Alford et al. 2010). Studies have found that AMF reduce (Liu et al. 2005; Jankong et al. 2007 ), increase (Wu et al. 2009; Orłowska et al. 2013), have mixed effects (Vogel-Mikuš et al. 2006), or no effects (Liu et al. 2009 ) on HMs uptake by hyperaccumulators. Up to date, only 4% of the over 700 reported hyperaccumulators have been studied in relation to AM symbiosis (Wang et al. 2021) , thus resulting in little understanding of the mechanisms involved in the interactions between plants, AMF and HMs (Dietterich et al. 2017). In nature, an equilibrium have been reached between AMF, host plants, and HMs over time (Zarei et al. 2008), therefore investigations of AMF communities at abandoned mining sites can greatly enhance the their utilization in phytoremedation (Sánchez-Castro et al. 2017 ).

Viola baoshanensis has been identified as a Cd hyperaccumulator, and also a strong accumulator of Zn and Pb by field survey at Baoshan Pb/Zn Mine, and by hydroponic and pot experiments ( Liu et al. 2004; Wu et al. 2010). This species could extract Cd and Zn with high efficiency in a field study (Zhuang et al. 2007) and could be well colonized by AMF in Cd and/or Pb contaminated soils in pot experiments (Zhong et al. 2012). However, the AM status of indigenous V. baoshanesis at the mining site remains unclear and need further research. The objectives of the present study were (1) to determine the accumulation of Cd, Zn and Pb by V. baoshanesis in relation to soil HMs concentrations; (2) to investigate the AM colonization status of V. baoshanesis in relation to soil chemo-physical properties ; (3) to examine the AMF community structure in the rhizosphere of V. baoshanesis.

2.1 Site description and sampling

Field sampling was conducted at Baoshan Pb/Zn Mine (25°42'N, 112°35'E), Guiyang County, Hunan Province, China, with a long history of mining and being abandoned in 1996 (Liu et al. 2004). Guiyang County has a humid subtropical monsoon climate with an annual average temperature and rainfall of 17.2°C and 1385 mm, respectively. V. baoshanensis is a perennial herb and endemic to the mine at an elevation of 400–650 m (Liu et al. 2004). Soil samples and plant material were collected at the flowering stage of V. baoshanensis in May, 2018. In total, 35 plants of V. baoshanensis were randomly collected at a slope near the mining pit, and soil samples were taken from the rhizosphere of the plants at a depth of 0–20 cm and stored at 4°C for further analysis.

2.2 Chemical analyses

The fresh soil samples were passed through 2-mm sieve and then divided into two parts. One portion was stored at 4°C at its real moisture status for the spore counting and identification, while the other was air-dried for the chemical analyses. The methods described by Zhong et al.(2012) were used to determine the total and DTPA extractable concentrations of Cd, Zn, and Pb of the soil samples, and total phosphorus of soil and plant samples. Soil pH was determined in a suspension (5:1, deionized water:soil), and measured by a pH electrode.

Plant shoots and roots were separated and about 1g fresh fine roots was left for observation of root AM colonization. The concentrations of Cd, Zn, and Pb of shoots and roots were determined by the methods described by Zhong et al.(2012). The characteristics of uptake and transfer of Cd, Zn and Pb by Viola baoshanensis were described by translocation factor (TF) and the bioconcentration factor (BF), which are defined by McGrath &Zhao (2003).

2.3 Root AM colonization

Fine roots of V. baoshanensis were cut into 1 cm segments cleared with 10% KOH and stained with 0.01% trypan blue ( Koske & Gemma 1989). 50 root segments per plant were examined under a compound microscope at 200× magnification to determine the mycorrhizal colonization by the magnified intersection method ( McGonigle et al. 1990). Mycorrhizal parameters such as mycorrhizal frequency (F%), relative mycorrhizal root length (M% ), relative arbuscular richness (A%), relative vesicular richness(V%) were used to assess the mycorrhizal status of V. baoshanensis.

2.4 Spore extraction and identification

Spores were isolated from soils by wet sieving and decanting followed by sucrose density gradient centrifugation (Tommerup & Kidby 1979 ). The extracted spores were mounted on the slides in polyvinyl alcohol-lactic acid glycerol (PVLG) and Melzer’s reagent for observation under a light microscope at 400-fold magnification. Spore identification was based mainly on morphological characteristics such as spore colour, wall structures, and subtending hyphae (Aguilera et al. 2014). Classification and taxonomic identification of AMF is according to identification reports (Redecker et al. 2013; Bills et al. 2015), species descriptions from INVAM homepage (http://www.invam.caf.wvu.edu), and other institutional collection of original species. Relative abundance of each species in one sample was determined by the percentage of the number of spores of the species in relation to the total number of spores in the sample ( Del Val et al. 1999).

2.5 Statistical analysis

Relative abundance of AMF species was tested for normality and analyzed by one-way analysis of variance (ANOVA), followed by the Duncan test at the level of 5% significance. Pearson’s correlation coefficient (r) was used when calculating correlations between mycorrhizal colonisation parameters, HMs concentrations of plants, and soil physico-chemical properties.

3.1 Soil chemical properties

The soil from the mining site was slightly alkaline (pH=7.60) and heavily polluted by Cd, Zn and Pb with maximal concentrations of 990 mgkg^-1Cd, 16548 mgkg^-1Zn, and 20111mgkg^-1 Pb. A significant variation in soil metal concentrations between 35 samples was observed as shown by large S.E.s (table 1), indicating the heterogeneous nature of the mining soil. Total and DTPA-extractable concentrations of Cd, Zn and Pb were shown in table1, while the total soil phosphorus was 857 ± 57 mgkg^-1.

Table 1 Cd, Zn and Pb concentrations of the soils from the rhizosphere of V. baoshanensis at Baoshan Pb/Zn Mine ( Mean ± S.E., n=35)

Total Metal (mgkg^-1)			DTPA-extractable metal (mgkg^-1)
Cd	Zn	Pb	Cd	Zn	Pb
147±30	7290 ± 720	7239 ± 694	36 ± 7	421 ± 42	506 ± 57

DTPA-extractable fractions ranged from 6.1 to 33.6% for Cd , from 0.9 to 9.7% for Zn, and from 4.5 to 13.1% for Pb. Significant correlations between total and DTPA-extractable concentrations were found for Cd ( r = 0.990, p < 0.01), Zn ( r = 0.767, p < 0.01 ), and Pb (r=0.871, p < 0.01), respectively. DTPA- extractable Cd, Zn, and Pb were 25%, 6% and 7 % of the total soil Cd, Zn and Pb, respectively. As the soil was slightly alkaline, it is reasonable that the bioavailability of these metals was low, which is in accordance with findings at the same site (Wu et al. 2010).

3.2 Cd, Zn , Pb and P accumulation in V. baoshanesis

As shown in table 2, the mean shoot Cd concentration was > 100 mgkg^-1, and the TF value was >1, thus confirming the hyperaccumulation of Cd by V. baoshanensis. Meanwhile, the BF value for Cd is 3.06, also showing high capacity of V. baoshanensis for the accumulation of Cd. Significant correlations were found between shoot Cd concentration and total Cd ( r=0.484, p<0.01), DTPA-extractable Cd (r=0.499, p<0.01) concentration in soil, and between root Cd concentration and total Cd (r=0.753, p<0.01), and DTPA-extractable Cd (r=0.747, p<0.01) concentration in soil, which is consistent with the results of Liu et al. (2004).

In contrast, no significant correlations were found in Zn concentrations between plants and soils. Although the TF of Zn was 2.57 indicating very efficient transport of Zn from roots to shoots, the shoot Zn concentration was much lower than the threshold, thus no hyperaccumulation of Zn was observed in V. baoshanensis, which was confirmed by the low BF value (average 0.15) as shown in table 2.

Table 2 Cd, Zn and Pb concentrations (mg kg⁻¹, DW) , BF and TF of V. baoshanensis collected at Baoshan Pb/Zn Mine ( Mean ± S.E., n=35 )

Metal	Shoot	Root	BF	TF
Cd	250±19	165±17	3.06±0.43	1.78±0.13
Zn	719±32	705±134	0.15±0.02	2.57±0.46
Pb	159±12	546±44	0.03±0.004	0.32±0.02

In contrast, the TF of Pb for V. baoshanensis was 0.32, indicating that most of the Pb accumulated was immobilized in roots. Shoot Pb concentration significantly correlated with total Pb ( r=0.398, p<0.05), DTPA-extractable Pb (r=0.361, p<0.05) concentration in soil, while root Pb concentration significantly correlated with total Pb (r=0.621, p<0.01), DTPA-extractble Pb (r=0.673, p<0.01 ) concentration in soil. The BF value for Pb was much lower than those for Cd and Zn. In addition, shoot and root P concentrations of V. baoshanensis were 1882 ± 93 and 4948 ± 357 mgkg^-1, respectively. Significant correlations were found between soil total P concentration and P concentrations in shoots or roots (r = 0.411, p < 0.05; r = 0.454, p < 0.01), respectively.

Accumulation and exclusion are two basic strategies taken by metallophytes growing on sites with high concentrations of HMs, the accumulation characteristics of which are commonly described by BF and TF values (McGrath &Zhao 2003). TF values over 1 in metal accumulator species are normal, whereas those in metal excluder species are often less than 1 (Baker 1981). In this survey, the shoot Cd concentration of V. baoshanensis exceeded the threshold, and the BF and TF for Cd were both greater than 1, thus confirming that V. baoshanensis is a Cd hyperaccumulator (Liu et al. 2004; Wu et al. 2010). The TF for Cd (1.78) of V. baoshanensis is comparable to that of a Cd, Zn hyperaccumulator, Sedum alfredii, naturally growing at a minging site with a TF of 1.87 (Yang et al. 2019). In addition, Thlaspi praecox, another Cd, Zn hyperaccumulator sampled from the vicinity of a lead mine and smelter in Slovenia, had a TF of 1.4 for Cd (Vogel-Mikuš et al. 2005). As hyperaccumulators are ideal candidates for phytoextraction, hyperaccumulation of Cd is of particular importance since so far only 10 species has been identified across the world (Wang et al. 2021).

As shoot concentration of Zn was lower than 10000 mg kg⁻¹, V. baoshanensis was not a Zn hyperaccumulator. However, the TF of V. baoshanensis for Zn was 2.57, suggesting there was efficient transfer of Zn from roots to shoots, while the TFs for Zn of S. alfredii and T. praecox were 1.24 and 3.2, respectively (Vogel-Mikuš et al. 2005; Yang et al. 2019). As for Pb, the TF is 0.32, indicating that most of Pb uptake were stored in roots. TFs of 0.3 were common for Pb in Thlaspi speices (Vogel-Mikuš et al. 2005), meanwhile the TF of S. alfredii for Pb was 0.28 (Yang et al. 2019).

3.3 Root colonization by AMF

High levels of colonization for V. basshanensis collected from the Baoshan Pb/Zn mine at flowering stage was observed with mean F% of 94.5%, and mean M% of 69.1% ranging from 51.3 to 83.8%. The roots were extensively colonized by intraradical hyphae forming well-developed arbuscules, but few vesicles were observed. The average A% was 46.9%, while the average V% was only 1.7% (Fig.1).

There were positive correlations between F% and M% (r = 0.628, P<0.01), between F% and A% (r = 0.401, P<0.05), and between M% and A% (r = 0.538, P<0.01). However, root colonization was not correlated with Cd, Zn and Pb concentrations in the soils, indicating that no inhibitory effect of HMs on mycorrhizal colonization of plant roots was evident.

Plant stage of development is one of the factors influencing AM colonization and flowering is a development reference point for the determination of AM status in calamine plants (Pawlowska et al. 1996), as the enhanced elemental demand during flowering favors mycorrhizal colonization, thus offering adequate essential elements (Pongrac et al. 2007) . Therefore, the plants of V. baoshanensis in this survey were collected at the flowering stage.

It has been suggested that selective pressure would decrease colonization unless the symbiosis supplied some benefits to the host (Whitfield et al. 2004). In contrast to several hyperaccumulators belonging to the genus Thlaspi poorly colonized by AMF (Regvar et al. 2003; Vogel-Mikuš et al. 2005), V. baoshanesis was well colonized by AMF. Colonization rate of 30% was considered as the threshold of a functional symbiosis (Zarei et al. 2010), the M% of V. baoshanesis was well above this level, indicating that AMF provide some benefits to their hosts in toxic soils (Whitfield et al. 2004; Rashid et al. 2009). Nevertheless, no significant correlation between the mycorrhizal status of V. baoshanesis and concentrations of metals in the soils was observed, which is consistent with the findings of Pierrier et al.(2006) . In addition, Viola tricolor L. grown on a calamine spoil mound (rich in Cd, Zn and Pb) was also found to be well colonized by AMF with percentage of root colonized of above 60% (Pawlowska et al. 1996), suggesting that the colonization status of a plant species may have a family basis.

As arbuscules are the major site of nutrient exchange (Smith & Read 1997), the arbuscular richness is considered as the most useful parameter for measuring mycorrhizal status ( Orłowska et al. 2005). A% of V. baoshanesis is considerably higher than those of Cd hyperaccumulators Desmostachya bipinnata and Dichanthium annulatum, which were 7% and 6%, respectively (Rashid et al. 2009), further indicating the existence of an effective symbiosis between V. baoshanesis and indigenous AMF.

3.4 AMF spore abundance and diversity

A total of 15 AMF species from 7 genera and 3 families of Glomeromycota were isolated and identified from the rhizosphere soil of V. baoshanensis (Table 3). Most of the species found belong to the family Glomeraceae (12 species ), and most of spores identified belonged to this family (Table 3). In terms of species richness, the most abundant genus was Glomus (5 speices ), and followed by the genus Rhizophagus (3 species) , while the relative abundance of Glomus was 44.0% , followed by Claroideglomus (18.5%) (Fig.2).

Table 3 Relative abundance (mean, n=35) of AM fungal species associated with Viola baoshanensis at Baoshan Pb/Zn Mine

AM Fungal species	Relative abundance (per cent)
Family Glomeraceae
Funneliformis mosseae	3.58 a,b,c
Glomus ambisporum	14.20 h
Glomus brohultii	6.44 b,c,d,e,f,g
Glomus formosanum	7.59 c,d,e,f,g
Glomus sp	9.80 f,g
Glomus hoi	6.00 a,b,c,d,e,f,g
Rhizophagus aggregatus	4.89 a,b,c,d,e
Rhizophagus fasciculatus	3.83 a,b,c,d
Rhizophagus invermiaum	1.82 a
Sclerocystis rubiformis	7.92 d,e,f,g
Septoglomus constrictum	8.86 e,f,g
Septoglomus deserticola	2.39 a,b
Family Claroideoglomeraceae
Claroideglomus claroideum	6.75 c,d,e,f,g
Claroideglomus etunicatum	11.79 g,h
Family Ambisporaceae
Ambispora leptoticha	4.17 a,b,c,d

Values followed by the same letters do not differ statistically by the Duncan test at the level of 5% significance.

There was a dominance of Glomeraceae species in HMs polluted areas such as minging tailings, which is a common observation worldwide (e.g. Zarei et al., 2010; Sun et al. 2016; Sánchez-Castro et al. 2017), showing that fungi in this family are more disturbance-tolerant than those in family Acaulosporaceae and Gigasporaceae (van der Heyde et al. 2017). In HMs polluted sites, soil pH has been found as an important factor in determining the AMF community structure ( Sun et al. 2016; Vieira et al. 2018; Yang et al. 2019). Acaulospora was widely observed in acid soils and Gigaspora are tolerant for acid pH (Clark 1997; van der Heyde et al. 2017), while species of Glomus are common in neutral and alkaline soils (da Silva et al. 2005). In consistent with this observation, the most abundant genus from the rhizosphere of V. banshanesis was Glomus, as the pH of the soil is slightly alkaline at Baoshan Pb/Zn mine. In addition, previous studies show that species from genus Claroideglomus are often tolerant to HMs (Cornejo et al. 2013; Park et al. 2016) and the results of this survey showed that Claroideglomus was the second abundant genus.

The number of identified spores was146 per 100 g soil on average. Glomus ambisproum and Claroideglomus etunicatum were the representative species, followed by Glomus sp and Septoglomus constrictum ( table 3) . Most of the AMF species identified in this survey have already been found in soils with high concentrations of HMs. Spores of G. ambisporum was found at a mining site contaminated with Zn, Cd, and Pb in southeast Kansas (USA), where S. constrictum was abundantly found (Shetty et al. 1994). C. etunicatum was identified from five of six sites in a copper minging area in northeastern Brazil (da Silva et al. 2005), meanwhile C. etunicatum was the most dominant species in the post-mining area (Korea), where S. constrictum and C. claroideum were also abundantly observed (Park et al. 2016). F. mosseae is normally found in soil contaminated with Zn and Pb (Turnau et al. 2001; Zarei et al. 2008), although the relative abundance of this species was only 3.58% in this study. The species identified in this survey have practical implications since metal-tolerant AMF should be isolated and inoculated in plants for phytoremediation (Vieira et al. 2019).

The proper integration of AMF and hyperaccumulators has been considered as the solution to pollution of HMs and finding appropriate AMF is very important (Vogel-Mikuš et al. 2006; Miransari M. 2011). The use of indigenous AMF species from long-term polluted sites has been widely observed as more suitable than laboratory strains (Sánchez-Castro et al. 2017 ). Up to now, information about interactions between native AMF and hyperaccumulators has been rather limited in published literature (Wang et al. 2021). Further work, therefore, need to focus on isolating AMF from the rhizosphere of native hyperaccumulators in the light of community structure to explore their roles in phytoremediation under laboratory and filed conditions.

Field survey confirmed that V. baoshanesis was a Cd hyperaccumulator but not a Zn hyperaccumulator, although efficient Zn transfer form roots to shoots was observed, while most of Pb uptake was stored in roots. This study showed that V. baoshanesis growing at Baoshan Pb/Zn mine with high concentrations of HMs were extensively colonized by AMF and forming well-developed arbuscules, suggesting that the host plant may derive some benefits from the symbiosis, and those AMF were HMs tolerant. Glomus was the dominant genus in the rhizoshpere of V. baoshanesis, while G. ambisporum and S. etunicatum was the most abundant species.

Al Agely A., Sylvia D. M., & Ma LQ. (2005). Mycorrhizae increase arsenic uptake by the hyperaccumulator Chinese brake fern (Pteris vittata L.). Journal of Environmental Quality, 34, 2181–2186. https://doi.org/10.2134/jeq2004.0411

Alford E. R., Pilon-Smits E. A.H., & Paschke M.W. (2010). Metallophytes – a view from the rhizosphere. Plant and Soil, 337, 337–350. https://doi.org/10.1007/s11104-010-0482-3

Antosiewicz D. M. , Escudĕ-Duran C., Wierzbowska E., & Skłodowska A. (2008). Indigenous plant species with the potential for the phytoremediation of arsenic and metals contaminated Soil. Water Air and Soil Pollution, 193, 197–210. https://doi.org/10.1007/s11270-008-9683-2

Arthur E. L., Rice P. J., Rice P. J., Anderson T. A., Baladi S. M., Henderson K. L. D., & Coats J. R. (2005). Phytoremediation – an overview. Critical reviews in plant sciences, 24, 109–122. https://doi.org/10.1080.07352680590952496

Audet P. (2013). Examining the ecological paradox of the ‘mycorrhizal- metal-hyperaccumulators’. Archives of Agronomy and Soil Science, 59, 549–558. https://doi.org/10.1080/03650340.2012.658378

Aguilera P., Cornejo P., Borie F., Barea J. M., von Baer E., & Oehl F. (2014). Diversity of arbuscular mycorrhizal fungi associated with Triticum aestivum L. plants growing in an Andosol with high aluminum level. Agriculture, Ecosystems and Environment, 186, 178–184. https://doi.org/10.1016/j.agee.2014.01.029

Baker A. J. M. (1981). Accumulators and excluders – strategies in the response of plants to heavy metals. Journal of Plant Nutrition, 3, 643–654.

Bills R. J., & Morton J. B. (2015). A combination of morphology and 28S rRNA gene sequences provide grouping and ranking criteria to merge eight into three Ambispora species (Ambisporaceae, Glomeromycota). Mycorrhiza, 25, 485–498. https://doi.org/10.1007/s00572-015-0626-7

Clark P. B. (1997). Arbuscular mycorrhizal adaptation, spore germination, root colonization, and host plant growth and mineral acquisition at low pH. Plant and Soil, 192, 15–22.

Cornejo P., Pérez-Tienda J., Meier S., Valderas A., Borie F., Azcón-Aguilar C., & Ferrol N. (2013). Copper compartmentalization in spores as a survival strategy of arbuscular mycorrhizal fungi in Cu-polluted environments. Soil Biology & Biochemistry, 57, 925–928. http://doi.org/10.1016/j.soilbio.2012.10.031

da Silva G. A., Trufem S. F. B., Júnior O. J. S., & Maia L. C. (2005). Arbuscular mycorrhizal fungi in a semiarid copper mining area in Brazil. Mycorrhiza, 15, 47–53. https://doi.org/10.1007/s00572-004-0293-6

Del Val C., Barea J. M., & Azcón-Aguilar C. (1999). Diversity of Arbuscular Mycorrhizal Fungus Populations in Heavy-Metal-Contaminated Soils. Applied and Environmental Microbiology, 65, 718–723.

Dietterich L. H.,Gonneau E., & Casper B. B. (2017). Arbuscular mycorrhizal colonization has little consequence for plant heavy metal uptake in contaminated field soils. Ecological Applications, 27, 1862–1875. https://doi.org/10.1002/eap.1573

Jankong P., Visoottiviseth P., & Khokiattiwong S. (2007). Enhanced phytoremediation of arsenic contaminated land. Chemosphere, 68, 1906–1912. https://doi.org/10.1016/j.chemosphere.2007.02.061

Khan A. G., Kuek C., Chaudhry T. M., Khoo C. S., & Hayes W. J. (2000). Role of plants, mycorrhizae and phytochelators in heavy metal contaminated land remediation. Chemosphere, 41, 197–207. https://doi.org/10.1016/S0045-6535(99)00412-9

Koske R. E. & Gemma J. N. (1989). A modified procedure for staining roots to detect VA mycorrhizas. Mycological Research, 92, 486–505.

Leyval C., Turnau K., & Haselwandter K. (1997). Effect of heavy metal pollution on mycorrhizal colonization and

function: physiological, ecological and applied aspects. Mycorrhiza, 7, 139–153.

Liu W., Shu W. S., & Lan C. Y. (2004). Viola baoshanensis, a plant that hyperaccumulates cadmium. Chinese Science Bulletin, 49, 29–32. https://doi.org/10.1360/03wc0245

Liu Y., Christie P., Zhang J., & Li X. (2009). Growth and arsenic uptake by Chinese brake fern inoculated with an arbuscular mycorrhizal fungus. Environmenal and Experimental Botany, 66, 435–441. http://doi.org/ 10.1016/j.envexpbot.2009.03.002

Liu Y., Zhu Y. G., Chen B.D., Christie P., & Li X. L. (2005). Influence of the arbuscular mycorrhizal fungus Glomus mosseae on uptake of arsenate by the As hyperaccumulator fern Pteris vittata L. Mycorrhiza, 15, 187–192. https://doi.org/10.1007/s00572-004-0320-7

McGonigle T. P., Miller M. H., Evans D. G., Fairchild G. L. & Swan J. A. (1990). A new method which gives an objective measure of colonization of roots by vesicular-arbuscular mycorrhial fungi. New Phytologist, 115, 495–501.

McGrath S.P., & Zhao F. (2003). Phytoextraction of metals and metalloids from contaminated soils. Current Opinion in Biotechnology, 14, 277–282. https://doi.org/10.1016/S0958-1669(03)00060-0

Miransari M. (2011). Hyperaccumulators, arbuscular mycorrhizal fungi and stress of heavy metals. Biotechnology Advances, 29, 645–653. https://doi.org/10.1016/j.biotechadv.2011.04.006

Orłowska E., Ryszka P., Jurkiewicz A., & Turnau K. (2005). Effectiveness of arbuscular mycorrhizal fungal (AMF) strains in colonisation of plants involved in phytostabilisation of zinc wastes. Geoderma, 129, 92–98. https://doi.org/10.1016/j.geoderma.2004.12.036

Orłowska E., Przybyłowicz W., Orlowski D., Mongwaketsi N. P., Turnau K. & Mesjasz-Przybyłowicz J. (2013). Mycorrhizal colonization affects the elemental distribution in roots of Ni-hyperaccumulator Berkheya coddii Roessler. Environmental Pollution, 175, 100–109. http://doi.org/10.1016/j.envpol.2012.12.028

Park H., Lee E., Ka K., & Eom A. (2016). Community Structures of Arbuscular Mycorrhizal Fungi in Soils and Plant Roots Inhabiting Abandoned Mines of Korea. Mycobiology, 44(4), 277–282. https://doi.org/10.5941/MYCO.2016.44.4.277

Pawlowska T. E., Błaszkowski J., & Rühling Å. (1996). The mycorrhizal status of plants colonizing a calamine spoil mound in southern Poland. Mycorrhiza, 6, 499–505.

Perrier N., Amir H., & Colin F. (2006). Occurrence of mycorrhizal symbioses in the metal-rich

lateritic soils of the Koniambo Massif, New Caledonia. Mycorrhiza, 16, 449–458. https://doi.org/10.1007/s00572-006-0057-6

Pongrac P., Vogel-Mikuš K., Kump P., Nečemer M., Tolrà R., Poschenriederet C., Barceló J., & Regvar M. (2007). Changes in elemental uptake and arbuscular mycorrhizal colonisation during the life cycle of Thlaspi praecox Wulfen. Chemosphere, 69, 1602–1609. https://doi.org/10.1016/j.chemosphere.2007.05.046

Rajkumar M., Sandhya S., Prasad M. N. V., & Freitas H. (2012). Perspectives of plant-associated microbes in heavy metal phytoremediation. Biotechnology Advances, 30, 1562–1574. https://doi.org/ 10.1016/j.biotechadv.2012.04.011

Redecker D., Schüßler A., Stockinger H., Stürmer S. L., Morton J. B., & Walker C. (2013). An evidence-based consensus for the classification of arbuscular mycorrhizal fungi (Glomeromycota). Mycorrhiza, 23, 515–531. https://doi.org/10.1007/s00572-013-0486-y

Regvar M., Vogel K., Irgel N., Wraber T., Hildebrandt U., Wilde P., & Bothe H. (2003). Colonization of pennycresses (Thlaspi spp.) of the Brassicaceae by arbuscular mycorrhizal fungi. Journal of Plant Physiology, 160, 615–626. https://doi.org/10.1078/0176-1617-00988

Sánchez-Castro I., Gianinazzi-Pearson V., Cleyet-Marel J. C., Baudoin E., & van Tuinen D. (2017). Glomeromycota communities survive extreme levels of metal toxicity in an orphan mining site. Science of the Total Environment, 598, 121–128. http://doi.org/10.1016/j.scitotenv.2017.04.084

Shetty K. G., Banks M. K., Hetrick B. A., & Schwab A. P. (1994). Biological characterization of a southeast Kansas mining site. Water, Air and Soil Pollution, 78, 169–177.

Smith S. E., & Read D. J. (1997). Mycorrhizal symbiosis. 2nd ed. Academic Press, San Diego, London.

Sun Y., Zhang X., Wu Z., Hu Y., Wu S., & Chen B. (2016). The molecular diversity of arbuscular mycorrhizal fungi in the arsenic mining impacted sites in Hunan Province of China. Journal of Environmental Sciences, 39, 110–118. http://doi.org/10.1016/j.jes.2015.10.005

Tommerup I. C. & Kidby D. K. (1979). Preservation of spores of vesicular arbuscular endophytes by L-drying. Applied

and Environmental Microbiology, 37, 831–835.

Turnau K., Ryszka P., Gianinazzi-Pearson V., & van Tuinen D. (2001). Identification of arbuscular mycorrhizal fungi in soils and roots of plants colonizing zinc wastes in southern Poland. Mycorrhiza, 10, 169–174.

Turnau K., & Mesjasz-Przybylowicz J. (2003). Arbuscular mycorrhiza of Berkheya coddii and other Ni-hyperaccumulating members of Asteraceae from ultramafic soils in South Africa. Mycorrhiza, 13, 185–190. http://doi.org/10.1007/s00572-002-0213-6

van der Heyde M., Ohsowski B., Abbott L. K., & Hart M. (2017). Arbuscular mycorrhizal fungus responses to disturbance are context-dependent. Mycorrhiza, 27, 431–440. http://doi.org/10.1007/s00572-016-0759-3

Vieira C. K., Marascalchi M. N., Rodrigues A. V., de Armas R. D., & Stürmer S. L. (2018). Morphological and molecular diversity of arbuscular mycorrhizal fungi in revegetated iron-mining site has the same magnitude of adjacent pristine ecosystems. Journal of Environmental Sciences, 67, 330–343. http://doi.org/10.1016/j.jes.2017.08.019

Vogel-Mikuš K., Drobne D., & Regvar M. (2005). Zn, Cd and Pb accumulation and arbuscular mycorrhizal colonisation of pennycress Thlaspi praecox Wulf. (Brassicaceae) from the vicinity of a lead mine and smelter in Slovenia. Environmental Pollution, 133, 233–242. http://doi.org/10.1016/j.envpol.2004.06.021

Vogel-Mikuš K., Pongrac P., Kump P., Nečemer M., & Regvar M. (2006). Colonisation of a Zn, Cd and Pb hyperaccumulator Thlaspi praecox Wulfen with indigenous arbuscular mycorrhizal fungal mixture induces changes in heavy metal and nutrient uptake. Environmental Pollution, 139, 362–371. http://doi.org/10.1016/j.envpol.2005.05.005

Wang L., Wang G., Ma F., & You Y. (2021). Symbiosis between hyperaccumulators and arbuscular mycorrhizal fungi and their synergistic effect on the absorption and accumulation of heavy metals: a review. Chinese Journal of Biotechnology. 37: 3604–3621. http://doi.org/10.13345/j.cjb.210305

Wei Y., Hou H., Li J., ShangGuan Y., Xu Y., Zhang J., Zhao L., & Wang W. (2014). Molecular diversity of arbuscular mycorrhizal fungi associated with an Mn hyperaccumulator - Phytolacca americana, in Mn mining area. Applied Soil Ecology, 82, 11–17. http://doi.org/10.1016/j.apsoil.2014.05.005

Whitfield L., Richards A. J., & Rimmer D. L. (2004). Relationships between soil heavy metal concentration and mycorrhizal colonisation in Thymus polytrichus in northern England. Mycorrhiza, 14, 55–62.

Wong M. H. (2003). Ecological restoration of mine degraded soils, with emphasis on metal contaminated soils. Chemosphere, 50, 775–780. https://doi.org/10.1016/S0045-6535(02)00232-1

Wu C., Liao B., Wang S., Zhang J. & Li J. (2010). Pb and Zn accumulation in a Cd-hyperaccumulator (Viola baoshanensis ). International Journal of Phytoremediation, 12, 574–585. http://doi.org/10.1080/15226510903353195

Wu F. Y., Ye Z. H., Wu S. C., & Wong M. H. (2007). Metal accumulation and arbuscular mycorrhizal status in metallicolous and nonmetallicolous populations of Pteris vittata L. and Sedum alfredii Hance. Planta, 226, 1363–1378. http://doi.org/10.1007/s00425-007-0575-2

Wu F. Y., Ye Z. H., & Wong M. H. (2009). Intraspecific differences of arbuscular mycorrhizal fungi in their impacts on arsenic accumulation by Pteris vittata L. Chemosphere, 76, 1258–1264. http://doi.org/10.1016/j.chemosphere.2009.05.020

Yang W., Li P., Rensing C., Ni W., & Xing S. (2019). Biomass, activity and structure of rhizosphere soil microbial community under different metallophytes in a mining site. Plant and Soil, 434, 245–262. https://doi.org/10.1007/s11104-017-3546-9

Zarei M., Saleh-Rastin N., Jouzani G. S., Savaghebi G. & Buscot F. (2008). Arbuscular mycorrhizal abundance in contaminated soils around a zinc and lead deposit. European Journal of Soil Biology, 44, 381–391. https://doi.org/10.1016/j.ejsobi.2008.06.004

Zarei M., Hempel S., Wubet T., Schäfer T., Savaghebi G., Jouzani G. S., Nekouei M. K. & Buscotet F. (2010). Molecular diversity of arbuscular mycorrhizal fungi in relation to soil chemical properties and heavy metal contamination. Environmental Pollution, 158, 2757–2765. https://doi.org/10.1016/j.envpol.2010.04.017

Rashid A., Ayub N., AHMsad T., Gul J., & Khan K. G. (2009). Phytoaccumulation prospects of cadmium and zinc by mycorrhizal plant species growing in industrially polluted soils. Environmental Geochemistry and Health, 31, 91–98. https://doi.org/10.1007/s10653-008-9159-8

Zhuang P., Yang Q.W., Wang H. B., & Shu W.S. (2007). Phytoextraction of heavy metals by eight plant species in the field. Water Air and Soil Pollution, 184, 235–242. https://doi.org/10.1007/s11270-007-9412-2

Zhong W., Li J., Chen Y., Shu W., & Liao B. (2012). A study on the effects of lead, cadmium and phosphorus on the lead and cadmium uptake efficacy of Viola baoshanensis inoculated with arbuscular mycorrhizal fungi. Journal of Environmental Monitoring, 14, 2497–2504. https://doi.org/10.1039/c2em30333g

Data availability statements

The data that support the findings of this study are not openly available due to sensitivity of the data and are available from the corresponding author upon reasonable request.

Statement of competing interests

The author declare no competing interests.

Funding declaration

No funding was received to assist with the preparation of this manuscript.

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Arbuscular mycorrhizal colonization of a Cd hyperaccumulator Viola baoshanensis at Baoshan Pb/Zn Mine

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Abstract

Figures

1. Introduction

2. Material And Methods

2.1 Site description and sampling

2.2 Chemical analyses

2.3 Root AM colonization

2.4 Spore extraction and identification

2.5 Statistical analysis

3. Results And Discussion

4. Conclusions

References

Declarations

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