Mikrobiol. Z. 2019; 81(4):42-53.
Detection of Phytopathogens Agrobacterium spp. and their Antagonists Bacillus thuringiensis,
Alcaligenes faecalis and Lactobacillus plantarum in Flowers and Berries of Grape
Limanska N., Galkin M., Marynova I., Ivanytsia V.
Odesa Mechnikov National University
2 Dvoryanskaya Str., Odesa, 65082, Ukraine
Aim. Detection of representatives of saprophytic (Bacillus thuringiensis, Alcaligenes faecalis, Lactobacillus plantarum) and pathogenic (Agrobacterium spp.) microbiota in flowers and berries of grape. Materials and Methods. Flowers, non-damaged green and ripe berries of grape were selected from a vineyard of Vitis vinifera L. cv Pinot noir located in Odessa region in May, July and September 2018, accordingly. Flowers or berries were homogenized and left for autofermentation process for 7 days. Real-Time polymerase chain reaction to detect species B. thuringiensis, L. plantarum, A. faecalis and Agrobacterium tumefaciens, Agrobacterium vitis was carried out with DNA isolated from the resulted fermented homogenate. Results. B. thuringiensis was prevalent among the tested species: these bacteria were identified in 50% of samples of grape flowers, in 90% of green berries and 20% of samples of ripe berries. L. plantarum was not detected in flowers, but these bacteria were identified in green and ripe berries (10% of tested samples). Results confirm literature data about the presence of B. thuringiensis and L. plantarum on grape. For the first time, we detected A. faecalis and Agrobacterium sрp. in flowers and berries of grape. A. faecalis was found in 20% of flower samples, 60% of green berries and 20% of ripe berries. Agrobacterium spp. were detected in flowers and green berries (10% of samples), and in ripe berries these microorganisms were not found. Amount of detected species increased in green berries compared with the flowers, and decreased in ripe berries. Conclusions. For the first time, we detected A. faecalis and Agrobacterium sрp. in flowers and berries of grape. Coexistence of phytopathogenic agrobacteria and their potential antagonists – Bacillus thuringiensis, Alcaligenes faecalis and Lactobacillus plantarum in one ecological niche (flowers and green grape berries) was revealed.
Keywords: Bacillus thuringiensis, Lactobacillus plantarum, Agrobacterium tumefaciens, Agrobacterium vitis, Alcaligenes faecalis, saprophytic microbiota, phytopathogens, Vitis vinifera.
Full text (PDF, in English)
- Bale JS, Lenteren van JC, Bigler F. Biological control and sustainable food production. Phil Trans R. Soc. B. 2008; 363:761-776. https://doi.org/10.1098/rstb.2007.2182
- Senna A de A, Lathrop A. Antifungal screening of bioprotective isolates against Botrytis cinerea, Fusarium pallidoroseum and Fusarium moniliforme. Fermentation. 2017; 53(3). https://doi.org/10.3390/fermentation3040053
- Bravo-Ferrada BM, Hollmann A, Delfederico L, Valdes La Hens D, Caballero A, Semorile L. Patagonian red wines: selection of Lactobacillus plantarum isolates as potential starter cultures for malolactic fermentation. World J Microbiol Biotechnol. 2013; 29(9):1537-1549. https://doi.org/10.1007/s11274-013-1337-x
- Behera SS, Ray SR, Zdolec N. Lactobacillus plantarum with functional properties: an approach to increase safety and shelf-life of fermented foods. BioMed Research International. 2018. https://doi.org/10.1155/2018/9361614
- Barata A, Malfeito-Ferreira M, Loureiro V. The microbial ecology of wine grape berries. Int J Food Microbiol. 2012; 153:243-259. https://doi.org/10.1016/j.ijfoodmicro.2011.11.025
- Bokulich NA, Collins TS, Masarweh C, Allen G, Heymann H, Ebeler SE, Mills DA. Associations among wine grape microbiome, metabolome, and fermentation behavior suggest microbial contribution to regional wine characteristics. mBio. 2016; 7(3):e00631-16. https://doi.org/10.1128/mBio.00631-16
- Mezzasalma V, Sandionigi A, Bruni I, Bruno A, Lovicu G, Casiraghi M, Labra M. Grape microbiome as a reliable and persistent signature of field origin and environmental conditions in Cannonau wine production. PloS ONE. 2017; 12(9): e0184615. https://doi.org/10.1371/journal.pone.0184615
- Shcherbakov AV, Mulina SA, Rots PYu, Shcherbakova EN, Chebotar VK. Bacterial endophytes of grapevine (Vitis vinifera L.) as promising tools in viticulture: isolation, characterization and detection in inoculated plants. Agronomy Research. 2016; 14(5):1702-1712.
- Bae SS. Investigation of bacteria associated with Australian wine grapes using cultural and molecular methods. Thesis for PhD degree. Sydney: University of New South Wales, School of Chemical Engineering and Industrial Chemistry; 2005.
- Kántor A, Kačániová M. Diversity of bacteria and yeasts on the surface of table grapes. Scientific Papers: Animal Science and Biotechnologies. 2015; 48:149-155.
- Maulani S, Hosseini SM, Elikaie A, Mirnurollahi SM. Isolated microorganisms from Iranian grapes and its derivatives. Iranian Journal of Microbiology. 2012; 4(1):25-29.
- Martins G, Lauga B, Miot-Sertier C, Mercier A, Lonvaud A, Soulas ML, Soulas G, Masneuf-Pomare I. Characterization of epiphytic bacterial communities from grapes, leaves, bark and soil of grapevine plants grown, and their relations. PLoS ONE. 2013; 8(8):e73013. https://doi.org/10.1371/journal.pone.0073013
- Pinto C, Pinho D, Sousa S, Pinheiro M, Egas C, Gomes A.C. Unravelling the diversity of grapevine microbiome. PLoS ONE 2014; 9(1)e85622. https://doi.org/10.1371/journal.pone.0085622
- Compant S, Mitter B, Colli-Mull JG, Gangl H, Sessitsch A. Endophytes of grapevine flowers, berries, and seeds: identification of cultivable bacteria, comparison with other plant parts, and visualization of niches of colonization. Microb Ecol. 2011; 62:188-197. https://doi.org/10.1007/s00248-011-9883-y
- Faist H, Keller A, Hentschel U, Deeken R. Grapevine (Vitis vinifera) crown galls host distinct microbiota. Appl Environ Microbiol. 2016; 82: 5542-5552. https://doi.org/10.1128/AEM.01131-16
- Kántor A, Mareèek J, Ivanisová E, Terentjeva M, Kacániová M. Microorganisms of grape berries. Proceedings of the Latvian Academy of Sciences. 2017; 71(6):502-508. https://doi.org/10.1515/prolas-2017-0087
- Bell CR, Dickey CA, Chan JWYF. Variable response of bacteria isolated from grapevine xylem to control grape crown gall disease in planta. American Journal of Enology and Viticulture. 1995; 46:499-508.
- Widnyana IK, Javandira C. Activities Pseudomonas spp. and Bacillus sp. to stimulate germination and seedling growth of tomato plants. Agriculture and Agricultural Science Procedia. 2016; 9:419-423. https://doi.org/10.1016/j.aaspro.2016.02.158
- Frikha-Gargouri O, Abdallah ben D, Ghorbel I, Charfeddine I, Jlaiel L, Triki MA, Tounsi S. Lipopeptides from a novel Bacillus methylotrophicus 39b strain suppress Agrobacterium crown gall tumours on tomato plants. Pest Manag Sci. 2017; 73(3):568-574. https://doi.org/10.1002/ps.4331
- Eastwell KC, Sholberg PL, Sayler RJ. Characterizing potential bacterial biocontrol agents for suppression of Rhizobium vitis, causal agent of crown gall disease in grapevine. Crop Protection. 2006; 25(11):1191-1200. https://doi.org/10.1016/j.cropro.2006.03.004
- Abdallah ben D, Frikha-Gargouri O, Tounsi S. Bacillus amyloliquefaciens strain 32a as a source of lipopeptides for biocontrol of Agrobacterium tumefaciens strains. Journal of Applied Microbiology. 2015; 119:196-207. https://doi.org/10.1111/jam.12797
- Banerjee G, Gorthi S, Chattopadhyay P. Beneficial effects of bio-controlling agent Bacillus cereus IB311 on the agricultural crop production and its biomass optimization through response surface methodology. Annals of the Brazilian Academy of Sciences. 2018; 90:2149-2159. https://doi.org/10.1590/0001-3765201720170362
- Bae S, Fleet GH, Heard GM. Lactic acid bacteria associated with wine grapes from several Australian vineyards. J Appl Microbiol. 2006; 100:712-727. https://doi.org/10.1111/j.1365-2672.2006.02890.x
- Sayyed RZ, Chincholkar SB. Siderophore-producing Alcaligenes feacalis exhibited more biocontrol potential vis-a'-vis chemical fungicide. Curr Microbiol. 2009; 58:47-51. https://doi.org/10.1007/s00284-008-9264-z
- Yokoyama S, Adachi Y, Asakura S, Kohyama E. Characterization of Alcaligenes faecalis strain AD15 indicating biocontrol activity against plant pathogens. J Gen Appl Microbiol. 2013; 59(2):89-95. https://doi.org/10.2323/jgam.59.089
- Kruchanova A, Korotaevа N, Limanskа N. Alcaligenes faecalis ONU 452 as an agent of bacterial biocontrol against Rhizobium radiobacter C58. In: Proceedings of the International Conference "Actual problems of Microbiology and Biotechnology"; 2015 June 1-4; Odessa Ukraine. Odessa: ONU Press, 2015. p. 18.
- Ultee A, Wacker A, Kunz D, Löwenstein R, König H. Microbial succession in spontaneously fermented grape must before, during and after stuck fermentation. S Afr J Enol Vitic. 2013; 34(1):68-78. https://doi.org/10.21548/34-1-1082
- Miranda-Castilleja DE., Martínez-Peniche RA, Aldrete-Tapia JA, Soto-Muñoz L, Iturriaga MH, Pacheco-Aguilar JR, Arvizu-Medrano SM. Distribution of native lactic acid bacteria in wineries of Queretaro, Mexico and their resistance to winelike conditions. Frontiers in Microbiology. 2016; 7:1769. https://doi.org/10.3389/fmicb.2016.01769
- Trias R, Bañeras L, Montesinos E, Badosa E. Lactic acid bacteria from fresh fruit and vegetables as biocontrol agents of phytopathogenic bacteria and fungi. Intern Microbiol. 2008; 11:231-236.
- Kharazian ZA, Jouzani GS, Aghdasi M, Khorvash M, Zamani M, Mohammadzadeh H. Biocontrol potential of Lactobacillus strains isolated from corn silages against some plant pathogenic fungi. Biological control. 2017. https://doi.org/10.1016/j.biocontrol.2017.04.004
- Limanska NV, Korotaeva NV, Yamborko GV, Ivanytsia VO. Effect of Lactobacillus plantarum on tumor formation caused by Rhizobium radiobacter. Microbiology and Biotechnology. 2014; 1:8-18. https://doi.org/10.18524/2307-4663.2014.1(25).48194
- Burr TJ, Bazzi C, Sule S, Otten L. Crown gall of grape: biology of Agrobacterium vitis and the development of disease control strategies. Plant Dis. 1998; 82:1288-1297. https://doi.org/10.1094/PDIS.1918.104.22.1688
- Burr TJ, Otten L. Crown gall of grape: biology and disease management. Annu Rev Phytopathol. 1999; 37:53-80. https://doi.org/10.1146/annurev.phyto.37.1.53
- Orel DC, Reid CL, Fuchs M, Burr TJ. Identifying environmental sources of Agrobacterium vitis in vineyards and wild grapevines. Am J Enol Vitic. 2017. https://doi.org/10.5344/ajev.2016.16085
- Ottesen AR, González Peña A, White JR, Pettengill JB, Li C, Allard S, Rideout S, Allard M, Hill T, Evans P, Strain E, Musser S, Knight R, Brown E. Baseline survey of the anatomical microbial ecology of an important food plant: Solanum lycopersicum (tomato). BMC Microbiol. 2013; 13:114. https://doi.org/10.1186/1471-2180-13-114
- Hassani A, Durán P, Hacquard S. Microbial interactions within the plant holobiont. Microbiome. 2018; 6:58. https://doi.org/10.1186/s40168-018-0445-0
- Pozo-Bayón MA, Pardo I, Sergi Ferrer S, Moreno-Arribas MV. Molecular approaches for the identification and characterisation of oenological lactic acid bacteria. African Journal of Biotechnology. 2009; 8(17):3995-4001.
- Torriani S, Felis GE, Dellaglio F. Differentiation of Lactobacillus plantarum, L. pentosus, and L. paraplantarum by recA gene sequence analysis and multiplex PCR assay with recA gene-derived primers. Appl Environm Microbiol. 2001; 67(8):3450-3454. https://doi.org/10.1128/AEM.67.8.3450-3454.2001
- Nakano M, Niwa M., Nishimura N. Development of a PCR-based method for monitoring the status of Alcaligenes species in the agricultural environment. Biocontrol Sci. 2014; 19:23-31. https://doi.org/10.4265/bio.19.23
- Park S-H, Kim HJ, Kim JH, Kim TW, Kim HY. Simultaneous detection and identification of Bacillus cereus group bacteria using multiplex PCR. J Microbiol Biotechnol. 2007; 17(7):1177-1182.
- Suzaki K, Yoshida K, Sawada H. Detection of tumorigenic Agrobacterium strains from infected apple saplings by colony PCR with improved PCR primers. J Gen Plant Pathol. 2004; 70:342-347. https://doi.org/10.1007/s10327-004-0133-8
- Bextine B, Lauzon C, Potter S, Lampe D, Miller TA. 2004. Delivery of a genetically marked Alcaligenes sp. to the glassy-winged sharpshooter for use in a paratransgenic control strategy. Curr Microbiol. 2004; 48(5):327-331. https://doi.org/10.1007/s00284-003-4178-2
- Liu J, Abdelfattah A, Norelli J, Burchard E, Schena L, Droby S, Wisniewski M. Apple endophytic microbiota of different rootstock/scion combinations suggests a genotypespecific influence. Microbiome. 2018; 6(1):18. https://doi.org/10.1186/s40168-018-0403-x
- Agrawal KS, Pathak RK. Phospate solubilization by Alcaligenes faecalis over Pseudomonas fluorescens. Agricultural Science Research Journals. 2012; 2:92-94.
- Limanska N, Ivanytsia T, Basiul O, Krylova K, Biscola V, Chobert J-M, Ivanytsia V, Haertle T. Effect of Lactobacillus plantarum on germination and growth of tomato seedlings. Acta Physiologiae Plantarum. 2013; 35:1587-1595. https://doi.org/10.1007/s11738-012-1200-y
- Aleklett K, Hart M, Shade A. The microbial ecology of ﬂowers: an emerging frontier in phyllosphere research. Botany. 2014; 92:253-266. https://doi.org/10.1139/cjb-2013-0166
- Shade A, McManus PS, Handelsman J. Unexpected diversity during community succession in the apple flower microbiome. MBio. 2013; 4(2):e00602-12. https://doi.org/10.1128/mBio.00602-12
- Mezzasalma V, Sandionigi A, Guzzetti L, Galimberti A, Grando MS, Tardaguila J, Labra M. Geographical and cultivar features differentiate grape microbiota in northern Italy and Spain vineyards. Frontiers in Microbiology. 2018; 9:946. https://doi.org/10.3389/fmicb.2018.00946
- Limanska N, Merlich A, Ivanytsia V. The spread of Lactobacillus plantarum in the fermented plant material from Ukraine and France. Visnyk of the Lviv University. 2016; 74:169-174.
- Groenewald WH, Reenen van CA, Dicks LMT. Strains of Lactobacillus plantarum in grape must are also present in the intestinal tract of vinegar flies. S Afr J Enol Vitic. 2006; 27(1):46-50. https://doi.org/10.21548/27-1-1599