Nitrogenase activity in in vitro associations between callus tissues of non-leguminous horticultural plants and rhizobium

1977 ◽  
Vol 9 (1) ◽  
pp. 1-5 ◽  
Author(s):  
Claus Schetter ◽  
Dieter Hess
2017 ◽  
Vol 9 (4) ◽  
pp. 27 ◽  
Author(s):  
Chie Shimaoka ◽  
Hirokazu Fukunaga ◽  
Seishu Inagaki ◽  
Shinichiro Sawa

The Orchidaceae are the largest and most diverse family of flowering plants on earth, and include some of the most important horticultural plants. While mycoheterotrophic orchids belonging to the genus Gastrodia are known to be provided with carbon through mycorrhizal fungi, the relationship between the plants and fungi is poorly understood. Furthermore, it is challenging to cultivate Gastrodia spp. in vitro. In this study, we present an efficient method for germinating Gastrodia pubilabiata (Gp), Gastrodia nipponica (Gn), and Gastrodia confusa (Gc) plants in vitro, which results in the production of a protocorm and tuber, as under natural conditions. The Gp and Gc plants produced flowers 126 and 124 days after germination, respectively, and set seed under our artificial conditions. In addition, Gp plants flowered up to three times a year from a single tuber. Using our artificial cultivation system, we identified some of the mycorrhizal fungi associated with these plants. Gastrodia spp. appear to obtain carbon from many kinds of mycorrhizal fungi. Our artificial cultivation method is a rapid and efficient means of growing Gastrodia spp. In addition to having applications in research and commercial nurseries, this method could be used to conserve Gastrodia spp. in ex situ, many of which are endangered.


2022 ◽  
Vol 8 ◽  
Author(s):  
Mohammad Imran Mir ◽  
Bee Hameeda ◽  
Humera Quadriya ◽  
B. Kiran Kumar ◽  
Noshin Ilyas ◽  
...  

A diverse group of rhizobacteria persists in the rhizospheric soil, on the surface of roots, or in association with rice plants. These bacteria colonize plant root systems, enhance plant growth and crop yield. Indigenous rhizobacteria are known to promote soil health, grain production quality and serve as sustainable bioinoculant. The present study was aimed to isolate, identify and characterize indigenous plant growth promoting (PGP) diazotrophic bacteria associated with the rhizosphere of rice fields from different areas of Jammu and Kashmir, India. A total of 15 bacteria were isolated and evaluated for various PGP traits, antagonistic activity against phytopathogens, production of hydrolytic enzymes and biofilm formation under in-vitro conditions. The majority of the isolated bacteria were Gram-negative. Out of 15 bacterial isolates, nine isolates produced IAA (12.24 ± 2.86 to 250.3 ± 1.15 μg/ml), 6 isolates exhibited phosphate solubilization activity (36.69 ± 1.63 to 312.4 ± 1.15 μg/ml), 7 isolates exhibited rock phosphate solubilization while 5 isolates solubilized zinc (10–18 mm), 7 isolates showed siderophore production, 8 isolates exhibited HCN production, 6 isolates exhibited aminocyclopropane-1-carboxylate (ACC) deaminase activity, 13 isolates exhibited cellulase activity, nine isolates exhibited amylase and lipase activity and six isolates exhibited chitinase activity. In addition, 5 isolates showed amplification with the nifH gene and showed a significant amount of nitrogenase activity in a range of 0.127–4.39 μmol C2H4/mg protein/h. Five isolates viz., IHK-1, IHK-3, IHK-13, IHK-15 and IHK-25 exhibited most PGP attributes and successfully limited the mycelial growth of Rhizoctonia solani and Fusarium oxysporum in-vitro. All the five bacterial isolates were identified based on morphological, biochemical and 16S rDNA gene sequencing study, as Stenotrophomonas maltophilia, Enterobacter sp., Bacillus sp., Ochrobactrum haematophilum and Pseudomonas aeruginosa. Rice plants developed from seeds inoculated with these PGP strains individually had considerably higher germination percentage, seed vigor index and total dry biomass when compared to control. These findings strongly imply that the PGP diazotrophic bacteria identified in this work could be employed as plant growth stimulators in rice.


2014 ◽  
Vol 69 (3) ◽  
pp. 193-195 ◽  
Author(s):  
Halina Ekiert ◽  
Wanda Kisiel

Four furanocoumarins: bergapten, xanthotoxin, isopimpinellin (linear furanocoumarins) and sphondin (angular furanocoumarin) were isolated for the first time from callus tissues of <em>Pastinaca sativa</em> L.(<em>Apiaceae</em>) cultured in vitro on solid medium. The compounds were identified using spectral methods. They are well-known secondary metabolites of the intact plant. This is the first report on the isolation of sphondin from in vitro plant cultures.


2016 ◽  
Vol 4 (1) ◽  
pp. 27 ◽  
Author(s):  
Edi Husen

Fourteen isolates of soil bacteria, including two known plant growth promoting rhizobacteria (PGPR) strains, Azotobacter vinelandii Mac 259 and Bacillus cereus UW 85, were tested in vitro. Parameters assessed were indoleacetic acid (IAA) production, phosphate solubilization, dinitrogen fixation, and siderophore (Fe-III chelating agent) production. IAA production was assayed colorimetrically using ferric chlorideperchloric acid reagent. Phosphate-solubilization and siderophore production were tested qualitatively by plating the bacteria in Pikovskaya and chrome azurol S agar, respectively. The ability to fix dinitrogen was measured based on nitrogenase activity of the bacteria by gas chromatography. The results showed that twelve isolates produced IAA, ranged from 2.09 to 33.28 µmol ml-1. The ability to solubilize precipitated phosphate was positively exhibited by four isolates (BS 58, BTS, TCaR 61, and BTCaRe 65). Seven isolates including Mac 259 positively produced siderophore. None of the isolates showed nitrogenase activity. Only one isolate (TS 3) did not exhibit any of the traits tested. Isolate TCeRe 60 and reference strain Mac 259 were found to have IAA- and siderophore-producing traits. Four P-solubilizing bacteria (BS 58, BTS, TCaR 61, and BTCaRe 65) were also IAA- and siderophore-producing bacteria. Potential use of these PGPR isolates needs further test in enhancing plant growth.


1981 ◽  
Vol 36 (1-2) ◽  
pp. 87-92 ◽  
Author(s):  
Patrick C. Hallenbeck ◽  
Leon V. Kochian ◽  
John R. Benemann

Abstract Cultures of Anabaena cylindrica, grown on media containing 5 mᴍ NH4Cl (which represses heterocyst formation), evolved hydrogen after a period of dark incubation under an argon atmosphere. This hydrogen production was not due to nitrogenase activity, which was nearly undetectable, but was due to a hydrogenase. Cultures grown on media with tungsten substituted for molybdenum had a high frequency of heterocysts (15%) and inactive nitrogenase after nitrogen starvation. The hydrogenase activity of these cultures was three-fold greater than the activity of non-heterocystous cultures. The effects of oxygen inhibition on hydrogen evolution by hetero-cystous cultures suggest that two pools of hydrogenase activity exist - an oxygen sensitive hydrogen evolution in vegetative cells and a relatively oxygen-resistent hydrogen evolution in heterocysts. In either case, inhibition by oxygen was reversible. Light had an inhibitory effect on net hydrogen evolution. Hydrogen production in vitro was much higher than in vivo, indicating that in vivo hydrogenase activity is limited by endogenous reductant supply.


Nitrogen ◽  
2020 ◽  
Vol 1 (2) ◽  
pp. 81-98
Author(s):  
Martina Lardi ◽  
Yilei Liu ◽  
Sebastian Hug ◽  
Samanta Bolzan de Campos ◽  
Leo Eberl ◽  
...  

Rhizobia have two major life styles, one as free-living bacteria in the soil, and the other as bacteroids within the root/stem nodules of host legumes where they convert atmospheric nitrogen into ammonia. In the soil, rhizobia have to cope with changing and sometimes stressful environmental conditions, such as nitrogen limitation. In the beta-rhizobial strain Paraburkholderia phymatum STM815, the alternative sigma factor σ54 (or RpoN) has recently been shown to control nitrogenase activity during symbiosis with Phaseolus vulgaris. In this study, we determined P. phymatum’s σ54 regulon under nitrogen-limited free-living conditions. Among the genes significantly downregulated in the absence of σ54, we found a C4-dicarboxylate carrier protein (Bphy_0225), a flagellar biosynthesis cluster (Bphy_2926-64), and one of the two type VI secretion systems (T6SS-b) present in the P. phymatum STM815 genome (Bphy_5978-97). A defined σ54 mutant was unable to grow on C4 dicarboxylates as sole carbon source and was less motile compared to the wild-type strain. Both defects could be complemented by introducing rpoNin trans. Using promoter reporter gene fusions, we also confirmed that the expression of the T6SS-b cluster is regulated by σ54. Accordingly, we show that σ54 affects in vitro competitiveness of P. phymatum STM815 against Paraburkholderia diazotrophica.


1980 ◽  
Vol 26 (9) ◽  
pp. 1072-1089 ◽  
Author(s):  
Dwight Baker ◽  
William Newcomb ◽  
John G. Torrey

The actinomycete, Frankia sp. EuI1, isolated from root nodules of Elaeagnus umbellata is an infective endophyte but which lacks the ability to form an effective nitrogen-fixing symbiosis with its host. This ineffective organism can be distinguished easily from other frankiae, in vitro, on the basis of size, morphology, and the elaboration of a diffusible pigment. Cross-inoculation studies indicated that the host range of this symbiont is narrow and probably restricted to the Elaeagnaceae. In all cases of nodulation the symbiosis never developed nitrogenase activity and the microsymbiont never produced endophytic vesicles within the infected host cells. Sporangia were produced in vivo and in vitro so the morphogenetic block is apparently restricted to vesicle formation.


1978 ◽  
Vol 24 (2) ◽  
pp. 143-148 ◽  
Author(s):  
J. J. Child ◽  
W. G. W. Kurz

Eleven different plant cell tissue cultures of both legume and non-legume origin have been grown in direct association, and in separate but close proximal association with both Spirillum lipoferum and Rhizobium sp. 32H1. Basic similarities were found in the nutritional requirement for the induction of nitrogenase activity (C2H2) in both organisms. In the absence of plant cell cultures both organisms need to be provided with a pentose sugar and a tricarboxylic acid to induce high levels of nitrogen-fixing activity. Plant cell callus tissue appears only capable of supplying the tricarboxylic acids needed but not the sugar component. The plant tissue, however, seems able to activate certain carbohydrates, which in themselves are incapable of substituting for the pentose additive.


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