Effect of GFP-tagging on nitrogen fixation and plant growth promotion of an endophytic diazotrophic strain of<i>Paenibacillus polymyxa</i>

Padda, Kiran Preet; Puri, Akshit; Zeng, Qingwei; Chanway, C. P.; Wu, Xiaoqin

doi:10.1139/cjb-2017-0056

Cited by 35 publications

(21 citation statements)

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“…This is in line with previous studies conducted on endophytes in mangrove plants found in Saudi Arabia that revealed the most dominant species being Bacillus [13]. Bacillus and related species have also shown presence of proteases, pectinases and cellulases in endophytic bacteria of Paenibacillus polymyxa [22]. The survival of these endophytes is attributed to their ability to utilize these cellulose and pectin in the plant tissues [22].…”

Section: Discussionsupporting

confidence: 90%

“…Bacillus and related species have also shown presence of proteases, pectinases and cellulases in endophytic bacteria of Paenibacillus polymyxa [22]. The survival of these endophytes is attributed to their ability to utilize these cellulose and pectin in the plant tissues [22]. The other dominant bacterial endophytes included the Streptomyces which was dominantly found in the leaves of Xylocarpus granatum plant.…”

Section: Discussionmentioning

confidence: 99%

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Enzymatic Activity of Endophytic Bacterial Isolates from Selected Mangrove Plants in Kenya

Ntabo¹,

Nyamache²,

Lwande³

et al. 2018

TOMICROJ

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Introduction:Microorganisms are a preferred source of enzyme production due to their high production capability and low cost of production. Bacterial endophytes occupy unexplored sites hence they represent a new source of enzymes with diverse applications. Mangrove plants in Kenya have traditionally been used as medicinal plants due to their bioactive metabolites. However the enzymatic activity of mangrove plants associated endophytes has not been studied.Aims & Objectives:The study is aimed at bioprospecting for enzymes with potential biotechnological applications from mangrove ecosystems.Methods & Materials:Forty-two bacterial isolates were cultured and isolated from the leaves and roots of six mangrove plants sampled from Mida Creek and Gazi Bay in the coastal region of Kenya. The isolates were screened for pectinases, chitinases, cellulases, proteases, and amylases. The isolates were identified based on morphology and 16S rRNA gene sequences analysis.Results:The study showed bacterial isolates had enzymatic activity as follows; pectinases activity (69% of the isolates), Proteases (95% of the isolates), amylases activity (88% of the isolates), cellulases and chitinases (92% of the isolates each). Bacterial endophytes from leaves showed a higher enzymatic index of cellulases suggesting a potential role in degrading cellulose in the leaves of plants. The enzymes amylases and proteases were mostly exhibited by endophytes in roots suggesting a potential role in metabolizing sugar and amino acids in the roots. Isolates from the mangrove plantSonneratia albashowed highest enzymatic indices. The study also observed that isolates from mangrove plants sampled from Gazi bay had high means of enzymatic indices. Molecular identification showed the isolates were closely related toBacillus, Streptomyces, Myroides, andStaphylococcusspecies. Their respective enzymatic activities have been provided in this study.Conclusion:The study showed that Kenyan Mangrove plant-associated bacterial endophytes provide a good reservoir of enzymes with potential industrial applications.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 99%

Enzymatic Activity of Endophytic Bacterial Isolates from Selected Mangrove Plants in Kenya

Ntabo¹,

Nyamache²,

Lwande³

et al. 2018

TOMICROJ

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“…Enhancement of these abilities in vitro after GFP-tagging were reported previously in Azospirillum brasilense [124]. A plausible reason for increased N fixing and plant growth-promoting efficacy of P2b-2R after GFP tagging could be the overexpression of structural nif genes (nifH, nifD, and nifK), which play an important role in the N-fixation process [121]. However, it is still unclear how GFP tagging affects the expression of structural nif genes of strain P2b-2R.…”

Section: Recent Studies Highlighting the Role Of Endophytic Diazotropmentioning

confidence: 53%

“…They tested this hypothesis by inoculating strain P2b-2R into agriculturally important crops, namely corn, canola, and tomato, and found that P2b-2R was able to colonize internal tissues of these crop species, fix substantial amounts of atmospheric N, and increase seedling length and biomass (see Table 2) [117][118][119]. These reports indicate the ability of strain P2b-2R to symbiotically associate with a broad range of hosts and promote their growth primarily by fixing atmospheric N. An interesting observation with the GFP-tagged P2b-2R strain (P2b-2Rgfp) was reported recently where P2b-2Rgfp inoculation significantly enhanced corn and canola seedling length and biomass as compared to the wild-type P2b-2R inoculation [119][120][121][122]. In addition, strain P2b-2Rgfp fixed significantly higher amounts of N as compared to the wildtype strain.…”

Section: Recent Studies Highlighting the Role Of Endophytic Diazotropmentioning

confidence: 61%

Nitrogen-Fixation by Endophytic Bacteria in Agricultural Crops: Recent Advances

Puri¹,

Padda²,

Chanway³

2018

Nitrogen in Agriculture - Updates

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Endophytic bacteria represents a unique class of bacteria that can colonize interior tissues of plant and provide a range of benefits to the plant similar to those provided by the rhizospheric bacteria. Certain endophytic bacteria can provide nitrogen to the plants through biological nitrogen fixation, which is an important source of nitrogen input in agriculture and represents a promising substitute for chemical fertilizers, and are known as endophytic diazotrophic bacteria. Besides fixing nitrogen, endophytic bacteria can produce plant growth hormones like auxin and gibberellin, help in nutrient uptake, and increase the plant's tolerance to biotic and abiotic stresses. Various direct and indirect methods have been used to quantify the amount of nitrogen fixed by these bacteria, including the acetylene reduction assay, which is a quick but indirect method, and the 15 N isotopic dilution assay, which is a robust and accurate method. Research on endophytic diazotrophic bacteria has come a long way, and in this chapter, we have briefly discussed the mechanisms of biological nitrogen fixation and methods to quantify the fixed nitrogen along with reviewing recent studies focused on evaluating the role of endophytic diazotrophic bacteria in promoting plant growth in both native and nonnative crop hosts.

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“…In fact, the endophyte Pseudomonas putida W619 presented a negative effect on poplar plants health and growth when GFP-labelled (Weyens et al, 2012). However, both the wild type and the GFP-tagged mutant of the diazotroph Paenibacillus polymyxa P2b-2R were able to promote the growth of pine (Tang et al, 2017), canola (Padda et al, 2016), and maize (Padda et al, 2017). In our work, a maize seed germination test was performed to ensure that the transformation did not affect the (which was not certified by peer review) is the author/funder.…”

Section: Discussionmentioning

confidence: 99%

Bacillus sp.RZ2MS9, a tropical PGPR, colonizes maize endophytically and alters the plant’s production of volatile organic compounds both independently and when co-inoculated withAzospirillum brasilenseAb-V5

Almeida

Bonatelli

Batista

et al. 2021

Preprint

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Bacillus spp. are among the most efficient known plant growth-promoting rhizobacteria (PGPR). The PGPR Bacillus sp. strain RZ2MS9 is a multi-trait maize growth promoter previously isolated from guarana plants cultivated in the Amazon rainforest. However, there are several aspects of its interaction with the host that need further investigation. To achieve effective performance of microbial inoculants in crop production, it is necessary to monitor the plant’s colonization by a PGPR and to assess the potential synergy among beneficial strains. Here, we obtained a stable mutant of RZ2MS9 labelled with green fluorescent protein (RZ2MS9-GFP). We verified that the insertion of the plasmid did not affect either bacterial growth nor its ability to promote maize growth in vitro. Using fluorescent microscopy and qPCR, we demonstrated that RZ2MS9-GFP successfully colonizes maize’s roots and leaves endophytically. Subsequently, we evaluated whether RZ2MS9 has a synergistic effect on plant growth promotion when co-inoculated with Azospirillum brasilense Ab-V5, a commercial inoculant for maize. The two strains combined enhanced maize’s roots and shoots dry weight by 50.8% and 79.6%, respectively, when compared to the non-inoculated control. In addition, we used co-inoculation experiments in glass chambers to analyze the plant’s Volatile Organic Compounds (VOCs) production during the maize-RZ2MS9 and maize-RZ2MS9-Ab-V5 interaction. We found that the single and co-inoculation altered maize’s VOCs emission profile, with an increase in the production of indoles in the co-inoculation. Collectively, these results increase our knowledge about the interaction between the tropical PGPR Bacillus sp. RZ2MS9 and maize, and provide a new possibility of combined application with the commercial inoculant A. brasilense Ab-V5.ImportanceBacillus sp. RZ2MS9 is a PGPR, previously isolated from guarana plants cultivated in the Brazilian Amazon, which successfully promotes the growth of maize and soybean plants. To improve our knowledge about the interaction between this very promising PGPR and maize, we labelled RZ2MS9 with gfp and monitored it’s maize colonization. The transformation did not affect either RZ2MS9 growth nor its ability to promote maize growth in vitro. We demonstrated that RZ2MS9 colonizes endophytically maize’s roots and leaves. We also verified that the co-inoculation of RZ2MS9 and Azospirillum brasilense Ab-V5, a known commercial maize inoculant enhanced maize’s roots and shoots growth. Moreover, the co-inoculation altered the maize’s volatile organic compounds, increasing the production of indoles, that is related with decreased upon the reduction of fertilization. Certainly, our research contributed with better Bacillus sp. RZ2MS9 – maize interaction understanding and also provided new information concerning RZ2MS9 activity when applied with A. brasilense Ab-V5.

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Effect of GFP-tagging on nitrogen fixation and plant growth promotion of an endophytic diazotrophic strain ofPaenibacillus polymyxa

Cited by 35 publications

References 49 publications

Enzymatic Activity of Endophytic Bacterial Isolates from Selected Mangrove Plants in Kenya

Enzymatic Activity of Endophytic Bacterial Isolates from Selected Mangrove Plants in Kenya

Nitrogen-Fixation by Endophytic Bacteria in Agricultural Crops: Recent Advances

Bacillus sp.RZ2MS9, a tropical PGPR, colonizes maize endophytically and alters the plant’s production of volatile organic compounds both independently and when co-inoculated withAzospirillum brasilenseAb-V5

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