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Arbuscular Mycorrhizal Colonization and Growth of Eremanthus Incanus Less

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Arbuscular Mycorrhizal Colonization and Growth of Eremanthus Incanus Less Arbuscular mycorrhizal colonization and growth of Eremanthus incanus Less. in a highland field M.C. Pagano, M.N. Cabello, M.R. Scotti Institute of Biological Sciences, Federal University of Minas Gerais, Belo Horizonte, Brazil ABSTRACT This paper focuses on Eremanthus incanus Less. (Asteraceae), a common species of highland regions in Brazil. The effect of arbuscular mycorrhizal (AM) inoculation on plant growth (height and diameter) was evaluated. Roots were examined from individuals randomly selected from undisturbed areas of highland vegetation and from an experimental restored site. Results showed that E. incanus presented high AM colonization both in restored and undisturbed sites. Moreover, AM colonization was significantly higher in the inoculated treatment than in the non- inoculated one. The species presented Arum-type colonization and frequent production of vesicles, especially in the restored site. Arbuscular mycorrhizal inoculation stimulated plant growth (height and diameter). Ten AM fungi (AMF) taxa were found in the studied rooting zones: Acaulospora spinosa, A. elegans, A. foveata, Acaulospora sp., Gigaspora margarita, Glomus sp., Dentiscutata biornata, D. cerradensis, Dentiscutata sp. and Racocetra verrucosa. These results revealed that AMF is a common and important component in highland vegetation in Brazil, and should be included in future restoration programs. Keywords: arbuscular mycorrhizal fungi; Eremanthus incanus; plant growth; highlands; Brazil A complete understanding of tropical plant life 2890 m, and its natural vegetation needs urgent histories should include traits related to AM for- strategies for conservation (Versieux and Wendt mation. Eremanthus incanus Less. (candeia, can- 2007). This fragile region has suffered from human deinha), common in southeast Brazil, in the States impact on a large scale, and conservation action of Minas Gerais and São Paulo, was selected to be must be developed to protect fauna and flora. studied. This arboreal species which reaches 3 to Highland fields, also termed rupestrian fields, have 5 m in height is common in Minas Gerais highland shrubby, tortuous and sclerophyllous vegetation or fields. This plant is presumed to be a threatened open grasslands and replace the cerrado (savanna) species and attempts to study its characteristics vegetation at 1000 m altitude. Plants grow in stones, are important for conservation and management in sandy soils and present varied adaptations (Giulietti because this species is subject to intense explora- and Pirani 1988). Typical transition forests composed tory pressure (Velten and Garcia 2007). predominantly of Eremanthus spp. (Asteraceae) are Arbuscular mycorrhizal fungi (AMF) are the main commonly found (Rizzini 1997). components of the microbial soil community, and One of the highland regions, the extreme south- their diversity is associated with plant community ern part of Espinhaço Range, is called ‘Quadrilátero structure and functioning; AMF diversity also Ferrífero’, due to exposed iron oxide deposits; it seems to be very important to determine produc- provides a habitat for many saxicolous species tivity and plant diversity in natural environments (Rizzini 1997). This is a region with high biodi- (Van der Heijden and Sanders 2003). AMF provide versity, presenting endemic and endangered spe- phosphate to their host plants, being particularly cies. Soils are shallow, acid, nutrient-poor, and important in soils with low P content (Bohrer et al. have excessively drained sands that are highly 2004); their sporulation is related to host species. erodable. The mycorrhizal associations of na- The State of Minas Gerais is characterized by tive species growing at highland fields have been a hilly relief with elevations ranging from 79 to scarcely studied. Nogueira et al. (2005) showed 412 PLANT SOIL ENVIRON., 56, 2010 (9): 412–418 the mycorrhizal status of some orchids; Pagano and replace the cerrado (savanna) vegetation from and Scotti (2009) showed AM colonization of two around 900–1000 m altitude (Giulietti and Pirani other species (Paepalanthus bromelioides and 1988). The climate is tropical montane (Aw type) Bulbostylis sp.); and Matias et al. (2009) reported with mean annual temperature varying between AM colonization for Centrosema coriaceum and 17 and 18.5°C, and mean annual precipitation estimated the spore numbers in the rhizosphere between 1450 and 1850 mm. Precipitation falls of C. coriaceum and Tibouchina multiflora, native primarily between December and March. species from this biome. This study was conducted at a highland field The main objective of this study was to investi- in restoration (43°26'W, 19°53'S) 1100 m a.s.l., in gate the effect of inoculation with AMF and the São Gonçalo do Rio Abaixo, Minas Gerais State, mycorrhizal associations of Eremanthus incanus and in an undisturbed adjacent site. Less. (Asteraceae), a native pioneer species in high- Treatments and experimental design. The ex- lands of southeastern Brazil. periment was set up in a degraded area used to keep Due to the urgent need for conservation of this iron-ore products. Eremanthus incanus plants were biome and the absence of any report for mycorrhizal cultivated in the experimental site (0.6 ha). Seedlings colonization of this plant species (Wang and Qiu were transplanted during the rainy season in 2001– 2006), this paper describes the AM associations 2002 and mixed with other 4 plant species. The of a native species to inform about conservation experimental design consisted of 12 replicate blocks and revegetation actions in the region. This is the (each 24 × 6 m). Two plants per plot randomized by first detailed report ever published on the AM species within the plot with 2-m spacing between status of this species. individual seedlings were used. Sixteen plants of E. incanus/block were used, thus mixed plantation in proportions of 2:10 (2 plants of E. incanus mixed MATERIALS AND METHODS with 10 plants of other 4 species/plot) was surveyed. The treatments were: 1 –E. incanus with com- Study area. The study area is located in the iron plete fertilization; 2 – E. incanus with complete mines region (43°30'W, 19°10'S), Minas Gerais fertilization (except for phosphorus) + inocula- State, in southeastern Brazil (Figure 1). The local tion with AMF. Complete fertilization consisted in landscape is highland fields, showing small woody P (218 kg/ha) using triple superphosphate, K (2 kg/ha) plants which support environmental stress (high as KCl, Mg (4.5 kg/ha) as MgSO4·7 H2O, Zn (13.6 kg/ day and night temperature oscillation and high ha) as ZnSO4·7 H2O, Mo (1.5 kg/ha), as Mo7O2·4 H2O, irradiance) (Rizzini 1997). Therefore, highly xero- urea (222 kg/ha) following Somasegaran and Hoben phytic vegetation with high plant species diversity (1985) and was applied at the beginning of the plan- and endemism is found (Rizzini 1997). These areas, tation. For inoculated treatment 50% of phosphorus termed rupestrian fields, have shrubby, tortuous fertilization was used. and sclerophyllous vegetation or open grasslands Arbuscular mycorrhizal fungi were inoculated by placing into each pot 1 ml of suspension composed by 100 spores/ml in a total of 3 species (Gigaspora margarita, Dentiscutata heterogama and Glomus etunicatum) using 33% of each species. Mycorrhizal fungi used were from the BHCB – UFMG collection. Site and soil characteristics. Soil samples were collected in 2003 from an undisturbed site in a highland field of southeastern Brazil, Minas Gerais State. Soil samples were kept in plastic bags, labeled, sealed and transported to the IMA (Instituto Mineiro de Agronomia) Agropecuary Chemical Laboratory (Brazil). Soils were air-dried and sieved with a sifter of 2 mm. Organic matter and phosphorus content (colorimetric method) was also determined. Potassium (K+), calcium (Ca2+), magnesium (Mg2+) and sodium (Na+) were deter- mined by atomic absorption spectrometry (Atomic Figure 1. Map showing the location of the sampling Absorption spectrophotometer 6800, Analytical site in the southern of Brazil PLANT SOIL ENVIRON., 56, 2010 (9): 412–418 413 Instruments Division Kyoto Japan, SHIMODZOU and thirty one-cm-root fragments were examined corporation, www.shimodzou.com). per sample for their AM status under a compound Soil texture was determined by the hydrom- microscope (100–1000 X). Quantification of my- eter method and soil pH was measured in H2O. corrhiza colonization was done noting the pres- Exchangeable K, Mg and Ca were determined by ence and absence of each of the fungal structures atomic-absorption spectrometry using 1N ammo- (intraradical hyphae and vesicles), and results were nium acetate as extracting solution. Exchangeable expressed as percentage of each structure in colo- Al was extracted with 1M KCl solution and deter- nized segments. Also the intensity of AMF colo- mined by titration with NaOH. nization was recorded (Pagano and Scotti 2009). Analysis of plant growth under field condi- These data were arcsin (x/100)1/2 transformed and tions. Plant height and diameter of E. incanus statistically analyzed, and means were compared were measured in field over 2 years (2002–2003), by the Tukey’s test (P < 0.05). within the dry and the rainy period. Growth data in both treatments were compared by the Tukey’s test (P < 0.05) following ANOVA. RESULTS AND DISCUSSION AM fungal distribution. Field work was carried out from March 2003 to December 2003, when The soil was sandy loam (0–30 cm depth). Some visits were conducted to collect rhizospheric soils basic
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