Skip to main content
×
×
Home

Exploration of methods used to describe bacterial communities in silage of maize (Zea mays) cultivars

  • Lorenzo Brusetti (a1), Sara Borin (a1), Aurora Rizzi (a1), Diego Mora (a1), Claudia Sorlini (a1) and Daniele Daffonchio (a1)...
Abstract

Different techniques to assess bacterial community structure and diversity were evaluated in silages prepared with four different maize cultivars, three conventional and one transgenic (cv. Tundra, event Bt-176). Plants were cultivated in the greenhouse and harvested after 30 days of growth. Silage samples were collected at successive times during fermentation and analyzed for bacterial counts and by various DNA-based fingerprinting techniques. Bacterial counts were similar between cultivars for the total culturable bacteria, sporeforming, and mesophilic and thermophilic lactic acid bacteria (LAB). Further analysis of the species composition of 388 LAB strains by intergenic transcribed spacer (ITS) PCR followed by sequencing of 16S rRNA gene did not reveal differences between cultivars. In contrast, molecular fingerprinting methods targeting whole bacterial communities, such as automated ribosomal intergenic spacers analysis (ARISA) and 16S rRNA gene length heterogeneity-PCR (LH-PCR), indicated that different maize silage batches or cultivars hosted different bacterial communities. Thus, ARISA and LH-PCR fingerprinting techniques offer a fast and sensitive method to compare bacterial communities, and to detect differences in silage bacterial communities.

    • Send article to Kindle

      To send this article to your Kindle, first ensure no-reply@cambridge.org is added to your Approved Personal Document E-mail List under your Personal Document Settings on the Manage Your Content and Devices page of your Amazon account. Then enter the ‘name’ part of your Kindle email address below. Find out more about sending to your Kindle. Find out more about sending to your Kindle.

      Note you can select to send to either the @free.kindle.com or @kindle.com variations. ‘@free.kindle.com’ emails are free but can only be sent to your device when it is connected to wi-fi. ‘@kindle.com’ emails can be delivered even when you are not connected to wi-fi, but note that service fees apply.

      Find out more about the Kindle Personal Document Service.

      Exploration of methods used to describe bacterial communities in silage of maize (Zea mays) cultivars
      Available formats
      ×
      Send article to Dropbox

      To send this article to your Dropbox account, please select one or more formats and confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your <service> account. Find out more about sending content to Dropbox.

      Exploration of methods used to describe bacterial communities in silage of maize (Zea mays) cultivars
      Available formats
      ×
      Send article to Google Drive

      To send this article to your Google Drive account, please select one or more formats and confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your <service> account. Find out more about sending content to Google Drive.

      Exploration of methods used to describe bacterial communities in silage of maize (Zea mays) cultivars
      Available formats
      ×
Copyright
Corresponding author
daniele.daffonchio@unimi.it
References
Hide All
[1] Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J. Mol. Biol. 215: 403–410
[2] Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1994) Current protocols in molecular biology. John Wiley and Sons, eds, USA
[3] Brookes G, Barfoot P (2006) GM Crops: The First Ten Years - Global Socio-Economic and Environmental Impacts. ISAAA Brief No. 36, ISAAA: Ithaca, NY
[4] Brusetti, L, Francia, P, Bertolini, C, Pagliuca, A, Borin, S, Sorlini, C, Abruzzese, A, Sacchi, G, Viti, C, Giovannetti, L, Giuntini, E, Bazzicalupo, M, Daffonchio, D (2004) Bacterial communities associated with the rhizosphere of transgenic Bt176 maize Zea mays and its non transgenic counterpart. Plant Soil 266: 1126
[5] Brusetti, L., Borin, S, Mora, D, Rizzi, A, Raddadi, N, Sorlini, C, Daffonchio, D (2006) Usefulness of length heterogeneity-PCR for monitoring lactic acid bacteria succession during maize ensiling. FEMS Microbiol. Ecol. 56: 154164
[6] Cai, Y, Benno, Y, Ogawa, M, Ohmomo, S, Kumai, S, Nakase T (1998) Influence of Lactobacillus spp. from an inoculant and of Weissella and Leuconostoc spp. from forage crops on silage fermentation. Appl. Environ. Microbiol. 64: 29822987
[7] Cai, Y, Kumai, S, Ogawa, M, Benno, Y, Nakase, T (1999) Characterization and identification of Pediococcus species isolated from forage crops and their application for silage preparation. Appl. Environ. Microbiol. 65: 29012906
[8] Cardinale, M, Brusetti, L, Quatrini, P, Borin, S, Puglia, AM, Rizzi, A, Zanardini, E, Sorlini, C, Corselli, C, Daffonchio, D (2004) Comparison of different primer sets for the Automated Ribosomal Intergenic Spacer Analysis (ARISA) of complex bacterial communities. Appl. Environ. Microbiol. 70: 61476156
[9] Chiarini, L, Bevivino, A, Dalmastri, C, Nacamulli, C, Tabacchioni, S (1998) Influence of plant development, cultivar and soil type on microbial colonisation of maize roots. Appl. Soil Ecol. 8: 1118
[10] Costa, R, Götz, M, Mrotzek, N, Lottmann, J, Berg, G, Smalla, K (2006) Effects of site and plant species on rhizosphere community structure as revealed by molecular analysis of microbial guilds. FEMS Microbiol. Ecol. 56: 236249
[11] Daffonchio, D, Borin, S, Frova, G, Manichini, PL, Sorlini, C (1998) PCR fingerprinting of whole genomes, the spacers between the 16S and 23S rRNA genes and of intergenic tRNA gene regions reveals a different intraspecific genomic variability of Bacillus cereus and Bacillus licheniformis. Int. J. Syst. Bacteriol. 48: 107116
[12] Di Giovanni, GD, Watrud, LS, Seidler, RJ, Widmer, F (1999) Comparison of parental and transgenic alfalfa rhizosphere bacterial communities using Biolog GN metabolic fingerprinting and enterobacterial repetitive intergenic consensus sequence-PCR (ERIC-PCR). Microb. Ecol. 37: 129139
[13] Dunfield, KE, Germida, JJ (2001) Diversity of bacterial communities in the rhizosphere and root interior of field-grown genetically modified Brassica napus. FEMS Microbiol. Ecol. 38: 19
[14] Escher, N, Käch, B, Nentwig, W (2000) Decomposition of transgenic Bacillus thuringiensis maize by microorganisms and woodlice Porcellio scaber (Crustacea: Isopoda). Basic Appl. Ecol. 1: 161169
[15] Fisher, MM, Triplett, EW (1999) Automated approach for ribosomal intergenic spacer analysis of microbial diversity and its application to freshwater bacterial communities. Appl. Environ. Microbiol. 65: 46304636
[16] Gomes, NCM, Heuer, H, Schönfeld, J, Costa, R, Medonça-Hagler, L, Smalla, K (2001) Bacterial diversity of the rhizosphere of maize (Zea mays) grown in tropical soil studied by temperature gradient gel electrophoresis. Plant Soil 232: 167180
[17] Griffiths, BS, Caul, S, Thompson, J, Birch, ANE, Scrimgeour, C, Andersen, MN, Cortet, J, Messé, A, Sausse, C, Lacroix, B, Krogh, PH (2005) A comparison of soil microbial community structure, protozoa and nematodes in field plots of conventional and genetically modified maize expressing the Bacillus thuringiensis CryIAb toxin. Plant Soil 275: 135146
[18] Gyamfi, S, Pfeifer, U, Stierschneider, M, Sessitsch, A (2002) Effects of transgenic gluphosinate-tolerant oilseed rape (Brassica napus) and the associated herbicide application on eubacterial and Pseudomonas communities in the rhizosphere. FEMS Microbiol. Ecol. 41: 181190
[19] Heuer, H, Kroppenstedt, RM, Lottmann, J, Berg, G, Smalla, K (2002) Effects of T4 lysozyme release from transgenic potato roots on bacterial rhizosphere communities are negligible relative to natural factors. Appl. Environ. Microbiol. 68: 13251335
[20] Koziel, MG, Beland, GL, Bowman, C, Carozzi, NB, Crenshaw, R, Crossland, L, Dawson, J, Desai, N, Hill, M, Kadwell, S, Launis, K, Lewis, K, Maddox, D, McPherson, K, Meghji, MR, Merlin, E, Rhodes, R, Warren, GW, Wright, M, Evola, SV (1993) Field performance of elite transgenic maize plants expressing an insecticidal protein derived from Bacillus thuringiensis. Bio-Technol. 11: 194200
[21] Lin, C, Bolsen, KK, Brent, BE, Hart, RA, Dickerson, JT, Feyerherm, AM, Aimutis, WR (1991) Epiphytic microflora on alfalfa and whole-plant corn. J. Dairy Sci. 75: 24842493
[22] Lin, C, Bolsen, KK, Brent, BE, Fung, DYC (1992) Epiphytic lactic acid bacteria succession during the pre-ensiling and ensiling periods of alfalfa and maize. J. Appl. Bacteriol. 73: 375387
[23] Nielsen, KM, Bones, AT, Smalla, K, van Elsas, JD (1998) Horizontal gene transfer from transgenic plants to terrestrial bacteria – a rare event? FEMS Microbiol. Rev. 22: 79103
[24] Ritchie, NJ, Schutter, ME, Dick, RP, Myrold, DD (2000) Use of Length Heterogeneity PCR and fatty acid methyl ester profiles to characterize microbial communities in soil. Appl. Environ. Microbiol. 66: 16681675
[25] Rizzi, A, Agosti, F, Daffonchio, D, Sorlini, C (2001) Detection of genetically modified Bt-maize in cooked food products by PCR. Ital. J. Food Sci. 13: 265274
[26] Rizzi, A, Panebianco, L, Giaccu, D, Sorlini, C, Daffonchio, D (2003) Stability and recovery of maize DNA during food processing. Ital. J. Food Sci. 15: 499510
[27] Saxena, D, Stotzky, G (2001) Bt corn has a higher lignin content than non-Bt corn. Am. J. Bot. 88: 17041706
[28] Saxena, D, Flores, S, Stotzky, G (1999) Insecticidal toxin in root exudates from Bt corn. Nature 402: 480
[29] Siciliano, SD, Germida, JJ (1999) Taxonomic diversity of bacteria associated with the roots of field-grown transgenic Brassica napus cv. Quest, compared to the non-transgenic B. napus cv. Excel and B. rapa cv. Parkland. FEMS Microbiol. Ecol. 29: 263272
[30] Smalla, K, Wieland, G, Buchner, A, Zock, A, Parzy, J, Kaiser, S, Roskot, N, Heuer, H, Berg, G (2001) Bulk and rhizosphere soil bacterial communities studied by denaturing gradient gel electrophoresis: plant-dependent enrichment and seasonal shifts revealed. Appl. Environ. Microbiol. 67: 47424751
[31] Smit, E, Leeflang, P, Gommans, S, van den Broek, J, van Mil, S, Wernars, K (2001) Diversity and seasonal fluctuations of the dominant members of the bacterial soil community in a wheat field as determined by cultivation and molecular methods. Appl. Environ. Microbiol. 67: 22842291
[32] Suzuki, M, Rappé, MS, Giovannoni, SJ (1998) Kinetic bias in estimates of coastal picoplankton community structure obtained by measurements of small-subunit rRNA gene PCR amplicon length heterogeneity. Appl. Environ. Microbiol. 64: 45224529
[33] Tukey, JW (1949) Comparing individual means in the analysis of variance. Biometrics 5: 99114
[34] Urzì, C, Brusetti, L, Salamone, P, Sorlini, C, Stackebrandt, E, Daffonchio, D (2001) Biodiversity of Geodermatophilaceae isolated from altered stones and monuments in the Mediterranean basin. Environ. Microbiol. 3: 471479
[35] Weinberg ZG, Muck RE (1996) New trends and opportunities in the development and use of inoculants for silage. FEMS Microbiol. Rev. 19: 53–68
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

Environmental Biosafety Research
  • ISSN: 1635-7922
  • EISSN: 1635-7930
  • URL: /core/journals/environmental-biosafety-research
Please enter your name
Please enter a valid email address
Who would you like to send this to? *
×

Keywords

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed