Bacteria associated with the roots of epiphytic orchids

This work is the first to report the isolation and identification of bacteria colonizing the roots of the tropical epiphytic orchids Acampe papillosa (Lindl.) Lindl. and Dendrobium moschatum (Buch.-Ham.) Swartz. and bacteria inhabiting inner layers of the aerial and substrate roots of A. papillosa. We showed by the example of this epiphyte that associative bacteria are present in large amounts on the aerial but not the substrate roots. We isolated and identified bacteria from the substrate roots of D. Moschatum and from its growth substrate (pine bark). The structure of the intercellular matrix of the associative bacteria was studied.Translated from Mikrobiologiya, Vol. 73, No. 6, 2004, pp. 825–831.Original Russian Text Copyright © 2004 by Tsavkelova, Cherdyntseva, Netrusov.     

Associative cyanobacteria isolated from the roots of epiphytic orchids

Associative cyanobacteria were isolated from the rhizoplane and velamen of the aerial roots of the epiphytic orchids Acampe papillosa, Phalaenopsis amabilis, and Dendrobium moschatum and from the substrate roots of Acampe papillosa and Dendrobium moschatum. Cyanobacteria were isolated on complete and nitrogen-free variants of BG-11 medium. On all media and in all samples, cyanobacteria of the genus Nostoc predominated. Nostoc, Anabaena, and Calothrix were isolated from the surface of the A. papillosa aerial roots, whereas the isolates from the substrate roots were Nostoc, Oscillatoria, and representatives of the LPP-group (Lyngbia, Phormidium, and Plectonema, incapable of nitrogen fixation). On the D. moschatum substrate roots, Nostoc and LPP-group representatives were also found, as well as Fischerella. On the aerial roots of P. amabilis and D. phalaenopsis grown in a greenhouse simulating the climate of moist tropical forest, cyanobacteria were represented by Nostoc, LPP-group, and Scytonema in the D. phalaenopsis and by Nostoc, Scytonema, Calothrix, Spirulina, Oscillatoria, and the LPP-group in P. amabilis. For D. moschatum, the spectra of cyanobacteria populating the substrate root zhizophane and the substrate (pine bark) were compared. In the parenchyma of the aerial roots of P. amabilis, fungal hyphae and/or their half-degraded remains were detected, which testifies to the presence of mycorrhizal fungi this plant. This phenomenon is attributed to the presence of a sheath formed by cyanobacteria and serving as a substrate for fungi.     

Localization of associative cyanobacteria on the roots of epiphytic orchids

This work is the first study of the localization of phototrophic microorganisms in the rhizoplane and velamen of epiphytic orchids, namely on the aerial and substrate roots of Acampe papillosa and Dendrobium moschatum and on the aerial roots of the Phalaenopsis amabilis and Dendrobium phalaenopsis. The composition of the bacterial community on the plant roots depended on the conditions of plant growth. Under conditions simulating climate of moist tropical forests, the aerial roots proved to be populated with phototrophic microorganisms among which cyanobacteria predominated. Interlaced fungal hyphae and filamentous cyanobacteria formed a sheath on the surface of aerial roots. The nitrogen-fixing capacity of the sheath of aerial roots was studied by the example of P. amabilis.     

Associative Cyanobacteria Isolated from the Roots of Epiphytic Orchids

Associative cyanobacteria were isolated from the rhizoplane and velamen of the aerial roots of the epiphytic orchids Acampe papillosa, Phalaenopsis amabilis, and Dendrobium moschatum and from the substrate roots of A. papillosa and D. moschatum. Cyanobacteria were isolated on complete and nitrogen-free variants of BG-11 medium. On all media and in all samples, cyanobacteria of the genus Nostoc predominated. Nostoc, Anabaena, and Calothrixwere isolated from the surface of the A. papillosa aerial roots, whereas the isolates from the substrate roots were Nostoc, Oscillatoria,and representatives of the LPP group (Lyngbia, Phormidium, and Plectonema, incapable of nitrogen fixation). On the D. moschatum substrate roots, Nostoc and LPP group representatives were also found, as well as Fischerella. On the aerial roots of P. amabilis and D. phalaenopsis grown in a greenhouse simulating the climate of moist tropical forest, cyanobacteria were represented by Nostoc, LPP group, and Scytonema in D. phalaenopsis and by Nostoc, Scytonema, Calothrix, Spirulina, Oscillatoria, and the LPP group in P. amabilis. For D. moschatum, the spectra of cyanobacteria populating the substrate root rhizoplane and the substrate (pine bark) were compared. In the parenchyma of the aerial roots of P. amabilis, fungal hyphae and/or their half-degraded remains were detected, which testifies to the presence of mycorrhizal fungi in this plant. This phenomenon is attributed to the presence of a sheath formed by cyanobacteria and serving as a substrate for fungi.     

Auxin production by bacteria associated with orchid roots

Bacteria associated with the roots of greenhouse tropical orchids were shown to produce indole-3-acetic acid (IAA) and to excrete it into the culture liquid. The presence and activity of IAA were demonstrated colorimetrically, by thin-layer chromatography, and by biotests. The associated bacteria varied in their ability to excrete indole compounds (1-28 microg/ml nutrient broth). Addition of tryptophan to the growth medium enhanced phytohormone production. Upon addition of 200 microg/ml tryptophan, the bacteria isolated from Dendrobium moschatum roots (Sphingomonas sp. 18, Microbacterium sp. 23, Mycobacterium sp. 1, Bacillus sp. 3, and Rhizobium sp. 5) produced 50.2, 53.1, 92.9, 37.6, and 60.4 microg IAA/ml respectively, while the bacteria isolated from Acampe papillosa roots (Sphingomonas sp. 42, Rhodococcus sp. 37, Cellulomonas sp. 23, Pseudomonas sp. 24, and Micrococcus luteus) produced 69.4, 49.6, 53.9, 31.0, and 39.2 microg IAA/ml. Auxin production depended on cultivation conditions and on the growth phase of the bacterial cultures. Treatment of kidney bean cuttings with bacterial culture liquid promoted formation of a "root brush" with location height 7.4- to 13.4-fold greater than the one in the control samples. The ability of IAA-producing associated bacteria to act as stimulants of the host plant root development is discussed.     

Microbiota of the Orchid rhizoplane

Six bacterial strains isolated from the underground roots of the terrestrial orchid Calanthe vestita var. rubrooculata were found to belong to the genera Arthrobacter, Bacillus, Mycobacterium, and Pseudomonas. Strains isolated from the aerial roots of the epiphytic orchid Dendrobium moschatum were classified into the genera Bacillus, Curtobacterium, Flavobacterium, Nocardia, Pseudomonas, Rhodococcus, and Xanthomonas. The rhizoplane of the terrestrial orchid was also populated by cyanobacteria of the genera Nostoc and Oscillatoria, whereas that of the epiphytic orchid was populated by one genus, Nostoc. In orchids occupying different econiches the spectra of the bacterial genera revealed differed. The microbial complex of the terrestrial orchid rhizoplane differed from that of the surrounding soil.     

Anti-diabetic potential of Barleria lupulina extract in rats.

We have undertaken a study to evaluate the anti-hyperglycemic effect of a methanol extract of aerial parts of Barleria lupulina Lindl. (Acanthaceae) in streptozotocin-diabetic rats, based on folkloric reports its use as an anti-diabetic agent. The extract exerted significant (p < 0.05) anti-hyperglycemic efficacy at all levels tested from 4 h after its administration, as compared with the control group, and the effect was also prolonged up to 12 h. The extract at doses of 200 mg kg(-1) body wt. and above exhibited a maximum activity (p < 0.001) at 12 h after administration. The most significant activity (15.35% blood glucose reduction) was observed for the group administered 300 mg kg(-1) body wt. at 12 h after administration, while the standard drug glibenclamide (10 mg/kg(-1) body wt.) showed an 18.80% reduction of blood glucose at the same time interval. Based on our current results, it appears that the methanol extract of aerial parts of Barleria lupulina Lindl. shows a pronounced blood-glucose-lowering potential in streptozotocin hyperglycemic rats, and is thus provided with a pharmacological support of the folklore claims of anti-diabetic activity.     

Resistance of red mangrove (Rhizophora mangle L.) seedlings to deflection and extraction

Red mangrove (Rhizhophora mangle L.) is the dominant tree species in the intertidal zone of ecosystems on the Atlantic shores of the Caribbean and tropical western Atlantic. The propagules of this species are initially buoyant, becoming negatively buoyant before rooting in a variety of substrates. After establishment, these seedlings form aerial roots, leading to communities of plants with complex networks of stems and aerial roots. While established mangrove communities assist in stabilizing coastlines, seedlings are susceptible to wave, current and wind energy and this limits the habitats that they can successfully colonize. In this experiment, the mechanical resistance of seedlings growing at five locations with different substrate and canopy conditions was tested. The 78 seedlings tested ranged in height from 27 to 47 cm, had between one and ten pairs of leaves but had not yet formed aerial roots. Seedlings were pulled horizontally. The reaction force at 20° deflection in four cardinal directions and then force to failure in the landward direction was measured. Seventy-five percent of the seedlings failed in the root system. The remainder failed near the base of the stem. Larger seedlings were more likely to fail at the roots. Seedlings growing outside of mangrove overstory on coral rubble were 3.5 times more strongly anchored than those growing within the mangrove overstory on sand. In spite of directional loading by waves and on-shore breezes, the deflection resistance did not vary systematically with pulling direction. Seedling anchorage varies among locations with different overstory and substrate conditions, likely due to differences in competition and acclimation to wind and wave energy along with differences in rooting among substrates.     

荔枝蝽生物学及防治技术的研究进展

荔枝蝽Tessaratoma papillasa是荔枝和龙眼生产中的重要害虫,在中国南方及东南亚地区危害严重.该虫以若虫及成虫吸食寄主植物的嫩芽、嫩梢、花穗和幼果,成虫和3龄以上若虫是传播龙眼鬼帚病病原体的传病介体昆虫.本文综述了近60年来国内荔枝蝽的研究情况,旨在为广大科研工作者提供更详尽的荔枝蝽研究现状,以期为该虫...     

Localization of Associative Cyanobacteria on the Roots of Epiphytic Orchids

This work is the first study of the localization of phototrophic microorganisms in the rhizoplane and velamen of epiphytic orchids, namely, on the aerial and substrate roots of Acampe papillosa and Dendrobium moschatum and on the aerial roots of Phalaenopsis amabilis and Dendrobium phalaenopsis. The composition of the bacterial community on the plant roots depended on the conditions of plant growth. Under conditions simulating the climate of moist tropical forests, the aerial roots proved to be populated with phototrophic microorganisms, among which cyanobacteria predominated. Interlaced fungal hyphae and filamentous cyanobacteria formed a sheath on the surface of the aerial roots. The nitrogen-fixing capacity of the sheath of the aerial roots was studied on the example of P. amabilis.     

Flavonoids and phenol carboxylic acids in the oriental medicinal plant Astragalus membranaceus acclimated in Poland

Astragalus membranaceus (Fisch.) Bunge has been was successfully acclimated in Central Europe. We report the content of isoflavones and some other polyphenolic compounds in roots and aerial parts that have been analyzed by means of TLC and HPLC. The total amount of isoflavones in leaves, was 0.55 mg g(-1) dry weight, and of the flavonols--up to 3.54 mg g(-1). In the roots isoflavonoid content was extremely variable, but reached 3.04 mg g(-1), whereas flavonols content was 0.49 mg g(-1).     

贵州兰科植物的新记录

报道了贵州兰科的3个新记录属:羽唇兰属、坛花兰属、异型兰属;以及9个新记录种:羽唇兰、坛花兰、异型兰、等萼卷瓣兰、广西舌喙兰、广西鸢尾兰、勐海隔距兰、长足石仙桃、长距虾脊兰。凭证标本保存在茂兰保护区标本室(HML)。     

青荚叶〔Helwingia japonica(Thunb.)Dietr.〕种下新组合

青荚叶〔Ｈｅｌｗｉｎｇｉａｊａｐｏｎｉｃａ (Ｔｈｕｎｂ .)Ｄｉｅｔｒ.〕为山茱萸科(Ｃｏｒｎａｃｅａｅ)青荚叶属 (ＨｅｌｗｉｎｇｉａＷｉｌｌｄ .)植物 ,我国民间常将其髓部、叶及果实作为药用。该种主要特征为叶片纸质 ,卵形、卵圆形、长卵形或卵状披针形 ,背面粉白或淡绿色等[1 ] 。作者在对馆藏的青荚叶标本进行整理时 ,发现《中国植物志》中的个别变种描述与该变种原始文献的描述有出入 ,而且该种种内的一些分类依据不明确。为此作者查阅了有关标本及文献 ,对该种的这些问题进行了分析和讨论。青荚叶Ｈｅｌｗｉｎｇｉａｊａｐｏｎ…     

Stemflow: A Source of Nutrients in some Naturally Growing Epiphytic Orchids of the Sikkim Himalaya

A study on five naturally growing epiphytic orchids viz., Bulbophyllum affine Lindl., Coelogyne ochracea Lindl., Otochilus porrecta Lindl., Cirrhopetalum cornutum Lindl. and C. cornutum (var.) was carried out in the subtropical belt of Sikkim Himalaya. Stemflow leachates formed the main source of ammonium-N and nitrate-N for uptake by these orchids. Phosphorus concentration in the tissues of these orchids was high. Phosphate-P from stemflow does not seem to be a regular source of phosphorus for these orchids. Absorption/desorption results indicate that organic-N from stemflow leachates is not utilized by these orchids.     

Effect of associative bacteria on element composition of barley seedlings grown in solution culture at toxic cadmium concentrations

The response of barley seedlings to inoculation with associative rhizobacteria Azospirillum lipoferum 137, Arthrobacter mysorens 7, Agrobacterium radiobacter 10 and Flavobacterium sp. L30 was studied in hydroponic and quartz sand cultures in the presence of 50 microM CdCl2. Cadmium caused severe inhibition in the growth and uptake of nutrient elements by the plants. Inoculation with the bacteria slightly stimulated root length and biomass of hydroponically grown Cd-treated seedlings. The bacteria increased the content of nutrients such as P, Mg, Ca, Fe, Mn and Na in roots and or shoots of the plants grown in the absence of Cd. Positive changes in the element composition caused by the bacteria were less pronounced in Cd-treated plants, whereas the total amount of nutrients taken by the inoculated plants was generally increased significantly. The content of Cd in the inoculated plants was unchanged, except increased in roots upon addition of A. lipoferum 137. Inoculation did not affect the activity of peroxidase, alpha-mannosidase, phosphodiesterae, alpha-galactosidase, and concentration of sulfhydryl compounds used as biochemical markers of stress in plant roots. The results showed that associative bacteria were capable of decreasing partially the toxicity of Cd for the barley plants through the improvement in uptake of nutrient elements.     

青荚叶[Helwingia japonica (Thunb.) Dietr.]种下新组合

Helwingia japonica (Thunb.) Dietr. var. papillosa Fang et Soong was reduced as a new synonym of H. japonica (Thunb.) Dietr. var. hypoleuca Hemsl. ex Rehd. Helwingia zhejiangensis Fang et Soong was combined into H. japonica (Thunb.) Dietr. and renamed H. japonica (Thunb.) Dietr. var. zhejiangensis (Fang et Soong) M. B. Deng et Y. Zhang.     

Enumeration and Localization of N(2)-Fixing Bacteria Associated with Roots of Spartina alterniflora Loisel

Numbers and possible locations of N(2)-fixing bacteria were investigated in roots of Spartina alterniflora Loisel, which support nitrogenase activity in the undisturbed native habitat. N(2)-fixing bacteria were recovered in cultures both from S. alterniflora roots and from the surrounding sediment, and they formed a greater proportion of the bacteria recovered from root homogenates than from salt-marsh sediment. N(2)-fixing bacteria were recovered in high numbers from the rhizoplane of S. alterniflora after roots were treated with 1 or 5% chloramine-T for 1 h or with 1% NaOCl for 1 or 2 h. Immersing S. alterniflora roots in 5% NaOCl for 1 h was more effective in distinguishing bacteria inside the roots since this treatment nearly eliminated N(2)-fixing bacteria recoverable from the rhizoplane, although high numbers of N(2)-fixing bacteria were recovered from homogenates of roots treated with 5% NaOCl for 1 h. However, this treatment was less effective with roots of Zea mays L. (Funks G4646) and Sorghum bicolor (L.) Moench (CK-60 A), indicating that techniques to surface sterilize roots should be evaluated for different plants. Bacteria were observed by light and electron microscopy inter- and intracellularly in the cortex and in the aerenchyma of S. alterniflora roots. This study clearly shows that bacteria, including N(2) fixers, colonize the interior of roots of S. alterniflora growing in a Chesapeake Bay, Maryland, salt marsh.     

Artificial colonization of non-symbiotic plants roots with the use of lectins

Rhizobia have the ability to increase growth of non-legume plants due to the production of phytohormones and protection of plant from diseases and pathogens. However, the practical use of these beneficial bacteria sometimes fails because of their inability to effectively colonize rhizoplane and rhizosphere of inoculated plants. We chose the legume lectins as a factor that allows plants to form associative symbiosis with rhizobia. To test the fact that transgenic tobacco, tomato and rape roots with pea lectin gene may affect specific interaction with rhizobia, transgenic roots have been artificially inoculated by fluorescently-labeled pea rhizobia R. leguminosarum and east galega rhizobia Rhizobium galega. Microscopic and microbiological tests have shown that the number of adhered R. leguminosarum onto tobacco, rape and tomato roots which transformed with pea lectin gene is higher in comparison with the control, but no such effect through inoculation of these plants with R. galegae has been found. This confirms the interaction of R. leguminosarum with pea lectin at the surface of transformed roots. Undoubtedly, the improvement of recognition and attachment processes by using lectins can lead to the achievement of a stable associative relationship between non-symbiotic plants and rhizobia.     

Growth of aerial roots with an extensive elongation zone by the example of a hemiepiphyte <Emphasis Type="Italic">Monstera deliciosa</Emphasis>

We investigated peculiarities of growth of aerial roots in a hemiepiphyte Monstera deliciosa. Aerial roots show low absolute and relative rates of growth and have an extensive elongation zone. In contrast to common roots, cell elongation in the elongation zone of aerial roots may last for 30 days and sometimes longer. The length of cortex cells increases in direct proportion to the distance from the root tip. This means that there is no drastic change in the relative rate of growth associated with transition to elongation characteristic of common roots. Distribution of growth over the elongation zone of aerial roots is irregular. Within the elongation zone, the cells of rhizodermis can divide, and divisions are distributed nonuniformly. The contact between neighboring growing polycytes (cell complexes) is presumably associated with their sliding against one another (intrusive growth). By the example of aerial roots of Monstera deliciosa, we showed a particular type of growth organization in the root with an extensive elongation zone differing from the growth of common roots and resembling the growth of leaves, stems, and fleshy fruit of dicotyledons.     

Root-associated n(2) fixation (acetylene reduction) by enterobacteriaceae and azospirillum strains in cold-climate spodosols

N(2) fixation by bacteria in associative symbiosis with washed roots of 13 Poaceae and 8 other noncultivated plant species in Finland was demonstrated by the acetylene reduction method. The roots most active in C(2)H(2) reduction were those of Agrostis stolonifera, Calamagrostis lanceolata, Elytrigia repens, and Phalaris arundinacea, which produced 538 to 1,510 nmol of C(2)H(4).g (dry weight). h when incubated at pO(2) 0.04 with sucrose (pH 6.5), and 70 to 269 nmol of C(2)H(4). g (dry weight).h without an added energy source and unbuffered. Azospirillum lipferum, Enterobacter agglomerans, Klebsiella pneumoniae, and a Pseudomonas sp. were the acetylene-reducing organisms isolated. The results demonstrate the presence of N(2)-fixing organisms in associative symbiosis with plant roots found in a northern climatic region in acidic soils ranging down to pH 4.0.