In vitro Decline in Plant Cultures: Detection of a Legion of Covert Bacteria as the Cause for Degeneration of Long-term Micropropagated Triploid Watermelon Cultures

In vitro cultures of triploid seedless watermelon cv. Arka Manik displayed a decline in shoot and root growth after 4–5 years of active culturing. Visibly clean cultures upon indexing on enriched media showed covert bacteria, and a significant improvement in proliferation and rooting in response to surface sterilization. The bacteria however survived endophytically. Low pH and reduced clarity of agar gelled medium were found to mask the expression of bacteria in the tissue culture medium. Gentamycin, streptomycin or broad-spectrum bactericide cefazolin provided as a 2 ml overlay in the medium in factorial combination at 0 or 50 mg l^–1 resulted in selective suppression of some bacteria depending on the treatment and eight bacterial clones comprising of four Gram-positive (Bacillus spp.) and four-Gram negative (3 × Pseudomonas spp. and 1 × Aeromonas sp.) strains were isolated from the cultures. Provision of 50 mg l^–1 gentamycin in 2 ml overlay in the multiplication or rooting medium coupled with occasional decontamination of cultures helped in circumventing the decline problem. The plants established in the field after 6 years of active in vitro culturing appeared normal and fertile suggesting the feasibility of keeping cultures for long periods, thus saving time and other resources. Freeing the cultures from covert bacteria was complicated by the presence of different bacterial types and this will be addressed later.     

内生真菌EPICOCCUM NIGRUM的形态与分子鉴定(英文)

在调查松属植物内生真菌过程中,从植物组织中分离到45株Epicoccum nigrum。根据它们在PDA培养基生长时的总体形态特征,划分为三组形态类型。从每组类型中随机选取三个代表菌株作进一步的分子鉴定。rDNA的ITS和5.8S基因序列分析结果表明,这9个菌株为E. nigrum。我们的研究结果也证明了前人提出的在Epicoccum菌种鉴定中,孢子大小和菌落颜色不能作为可靠的种级分类特征。     

植物内生菌抗虫工程研究进展

生物防治已成为植物虫害防治的发展方向,越来越多地受到人们的关注。利用植物内生菌进行抗虫性的研究不断深入。本着重论述了内生菌的抗虫机制及其影响因素。同时,对抗虫策略的实施也进行了探讨。     

Identification of the endophypte ofDryas andRubus (Rosaceae)

Summary Root nodules ofDryas drummondii are of the coralloid type (Alnus type). The endophyte is present in the middle cortical cells of the root-nodule tissue. Transmission electron micrographs revealed an actinorhizal endophyte with septate hyphae and non-septate spherical or ovoid vesicles. Vesicles always possess at the base a septum; septa formation in the endophyte is always associated with the presence of mesosomes. Branching of the endophyte is not necessarily correlated with septum formation. Hyphal structures are more prominent in the apical part of the root nodule and vesicles are more numerous in a broad zone below this. In the middle and towards the base of the root nodule the endophytic structures appear in a stage of disintegration. Vesicles appear in a broad region near the periphery of the host cell and regularly show no strict orientation towards the host-cell wall. In the center of the host cells only hyphae occur. In the intercellular spaces between the host cells theFrankia endophyte produces spore-like structures although the outline of the sporangia is often faint.The coralloid root ofRubus ellipticus shows characteristically a basal rootlet initiated below the dichotomous branching of the nodular lobes, but extending beyond the root nodule. The endophyte is only present in the outer cortex of the root nodule in a 1–2 cell wide layer. This endophytic layer is bounded, internally as well as externally, with a 4–5 cell wide layer of tannin-filled host cells. The implications of this situation are discussed. Tannin-filled cells occur regularly inRubus species and their arrangement has been used for taxonomic purposes within the genus. TheRubus endophyte is aFrankia species with septate hyphae and distinctly septate spherical vesicles. The ultrastructure of the vesicles of theRubus endophyte is very similar to that of theAlnus endophyte.     

Ammonia cometabolism and product inhibition vary considerably among species of methanotrophic bacteria

The structural gene for 1-aminocyclopropane-1-carboxylate (ACC) deaminase ( acdS ) from the endophytic plant growth-promoting bacterium Burkholderia phytofirmans PsJN was isolated and used to construct a mutant strain B. phytofirmans YS2 ( B. phytofirmans PsJN/Δ acdS ), in which an internal segment of the acdS gene was deleted. The mutant YS2 lost ACC deaminase activity as well as the ability to promote the elongation of the roots of canola seedlings. Concomitant with the creation of this deletion mutant, a number of physiological changes were observed in the bacterium, including an increase in indole acetic acid synthesis, a decrease in the production of siderophores and an increase in the cellular level of the stationary-phase σ factor, RpoS. Introduction of the wild-type acdS gene into the mutant YS2 to construct strain B. phytofirmans YS3 ( B. phytofirmans YS2/pRK-AcdS) restored both ACC deaminase activity and plant growth-promotion activity in strain YS3. However, the complemented mutant still showed the above-mentioned physiological changes.     

植物内生菌在生物防治中的应用

从植物内生菌的抗病虫机理及生防作用方面综述了植物内生菌在生物防治中的研究进展,对植物内生菌的抗病虫机理进行了探讨,并展望了植物内生菌在植物保护中的应用前景.     

滑桃树内生真菌Fusarium sp.1RGa-1b中萘醌类代谢产物及其抗菌活性研究

从滑桃树内生真菌Fusarium sp．（1RGa—1b）的发酵代谢产物中分离得到6个一系列的萘醌类色素化合物,通过波谱分析将其结构鉴定为：3-methylether-fusarubin（1）,anhydrofusarubin（2）,2-aeetonyl-3-methyl-5-hydro—gen-7-methoxy—naphthazarin（3）,2-acetonyl-3-methyl-7-methoxy-8-hydrogen—naphthazarin（4）,2-acetonyl-3-methyl-7-methoxy—naphthazarin（5）,2-isopropanol-3-methyl-7-methoxy-naphthazafin（6）。利用纸片扩散法对这些化合物的抗细菌及抗真菌活性进行了初步测试,实验结果表明：化合物3和4具有弱的抗金黄色葡萄球菌的活性。     

四个品种烟草内生微生物的分布特征

对4个常规优质烟草品种(K326、G-80、红花大金元、净叶黄)的种子、根、茎、叶进行了内生菌的分离计数、初步分类与拮抗筛选研究.结果表明:烟草中有大量的内生菌分布,以细菌最多,放线菌次之,真菌最少,与土壤中的分布规律一致.烟草不同器官中内生菌的分布表现为根和种子中数量相当,茎次之,叶最少的规律.不同烟草品种间的分布为:种子中是K326和红花大金元内生菌分布较多,净叶黄次之,G-80最少;苗期植株中4个品种的内生菌数没有明显差别;大田期是净叶黄烟草的内生菌数量最少.烟草中内生细菌以革兰氏阴性菌为主,而种子中内生细菌的芽胞菌数量要明显多于植株中的.通过对大肠杆菌和金黄色葡萄球菌的抑菌实验,从红花大金元品种中筛选到有抑菌活性的菌株5株,从净叶黄和K326中分别筛选到1株,体现出了4个品种的抗病性与拮抗菌之间的一定相关性.     

Trichoderma species form endophytic associations within Theobroma cacao trichomes

Trichoderma species are usually considered soil organisms that colonize plant roots, sometimes forming a symbiotic relationship. Recent studies demonstrate that Trichoderma species are also capable of colonizing the above ground tissues of Theobroma cacao (cacao) in what has been characterized as an endophytic relationship. Trichoderma species can be re-isolated from surface sterilized cacao stem tissue, including the bark and xylem, the apical meristem, and to a lesser degree from leaves. SEM analysis of cacao stems colonized by strains of four Trichoderma species (Trichoderma ovalisporum-DIS 70a, Trichoderma hamatum-DIS 219b, Trichoderma koningiopsis-DIS 172ai, or Trichoderma harzianum-DIS 219f) showed a preference for surface colonization of glandular trichomes versus non-glandular trichomes. The Trichoderma strains colonized the glandular trichome tips and formed swellings resembling appresoria. Hyphae were observed emerging from the glandular trichomes on surface sterilized stems from cacao seedlings that had been inoculated with each of the four Trichoderma strains. Fungal hyphae were observed under the microscope emerging from the trichomes as soon as 6 h after their isolation from surface sterilized cacao seedling stems. Hyphae were also observed, in some cases, emerging from stalk cells opposite the trichome head. Repeated single trichome/hyphae isolations verified that the emerging hyphae were the Trichoderma strains with which the cacao seedlings had been inoculated. Strains of four Trichoderma species were able to enter glandular trichomes during the colonization of cacao stems where they survived surface sterilization and could be re-isolated. The penetration of cacao trichomes may provide the entry point for Trichoderma species into the cacao stem allowing systemic colonization of this tissue.     

<Emphasis Type="Italic">Heteroconium chaetospira</Emphasis>, a dark septate root endophyte allied to the Herpotrichiellaceae (Chaetothyriales) obtained from some forest soil samples in Canada using bait plants

During an extended search in Western Canada for fungal root endophytes useful as biocontrol agents against soil-borne pathogens, we isolated Heteroconium chaetospira, as well as Phialocephala fortinii or similar taxa, from seven samples of forest soil using herbaceous seedlings of four different species (i.e., barley, Chinese cabbage, eggplant, and melon) as bait plants. Our results support a previous observation that eggplant is a particularly effective species for baiting H. chaetospira from soil and confirm the ability of this fungus to grow as an endophyte in the roots of axenically reared host plants. Cultural characters show that this species is similar to P. fortinii and other melanized fungi in the dark septate endophyte (DSE) group (e.g., Leptodontidium orchidicola, P. sphaeroides, and Cadophora finlandica) in that it produces darkly pigmented colonies on agar media. Heteroconium chaetospira differs from P. fortinii and other melanized members of the Leotiomycetes in the DSE group in that its conidia are fusiform and develop in blastic acropetal chains. Heteroconium chaetospira is phylogenetically distant from most DSE taxa because DNA sequences for the nuclear small subunit (SSU) ribosomal RNA gene (rDNA) indicate that the taxon is affiliated with the Herpotrichiellaceae of the Chaetothyriales rather than with the Leotiomycetes.