scholarly journals Occurrence of Phytoplasma Belonging to 16SrII Group Associated with Witches’-broom Symptoms in Emilia sonchifolia in Hainan Island of China

Plant Disease ◽  
2021 ◽  
Author(s):  
Shao-shuai Yu ◽  
Rui-ling Zhao ◽  
Ming-xing Lin ◽  
Yuan Wu ◽  
Chen Shu-gui ◽  
...  

Emilia sonchifolia is a medical plant belonging to the family of Asteraceae, mainly used as a traditional Chinese medicine with the function of anti-inflammatory, analgesic, antibacterial and so on. During October to November 2020, the plants showing abnormal symptoms including witches’-broom, internode shortening, leaf chlorosis and leaflet were found in Hainan province, a tropical island of China. The total DNA of the plant samples were extracted using 0.10 g fresh plant leaves using CTAB method. PCR reactions were performed using primers R16mF2/R16mR1 and secAfor1/secArev3 specific for phytoplasma 16S rRNA and secA gene fragments. The target productions of the two gene fragments of phytoplasma were detected in the DNA from three symptomatic plant samples whereas not in the DNA from the symptomless plant samples. The two gene fragments of the DNA extracted from the symptomatic plant samples were all identical, with the length of 1324 bp 16S rRNA and 760 bp secA gene sequence fragments, putatively encoding 253 (secA) amino acids sequence. The phytoplasma strain was named as Emilia sonchifolia witches’-broom (EsWB) phytoplasma, EsWB-hnda strain. To our knowledge, this was the first report that Emilia sonchifolia witches’-broom disease was caused by the phytoplasma belonging to16SrII-V subgroup in Hainan island of China, with close relationship to 16SrII peanut witches’-broom group phytoplasma strains infecting the plants like peanut, Desmodium ovalifolium and cleome from the same island of China and cassava from Viet Nam.

Plant Disease ◽  
2020 ◽  
Author(s):  
Shao-shuai Yu ◽  
Qinghua Tang ◽  
Yuan Wu ◽  
Rui-ling Zhao ◽  
Wei wei Song ◽  
...  

Pericampylus glaucus is an important medicinal plant resource containing active components with potential antitumor activity in China (Zhao & Cui, 2009). During July through August 2020, plants displayed disease symptoms including “witches’ broom”, leaf chlorosis, leaflet and internode shortening that impacted their growth (Fig. 1). These plants were first found in Dingan county of Hainan province, China. Total DNA from 12 plants were extracted using 0.10 g fresh plant leaves based on CTAB method. After amplification using primers specific for phytoplasma 16S rRNA, tuf and secA gene targets, R16mF2R16mR1 (Lee et al, 1993), fTuf1/rTuf1 (Schneider et al., 1997) and secAfor1/secArev3 (Hodgetts et al., 2008), the target bands of the three gene fragments of phytoplasma were detected in the disease sample DNA from six disease plants, and not in the healthy sample DNA from six healthy plants. Nucleotide sequences of the three genes were obtained from the PCR products sequencing and analyzed by DNAMAN 5.0 software. The three gene fragments of the DNA extracted from the disease samples were identical, with length of 1334 bp 16S rRNA (GenBank accession: MT872515), 989 bp tuf (MT755960) and 750 bp secA (MT755961) gene fragments, putatively encoding 329 (tuf) and 249 (secA) amino acids sequence separately. The phytoplasma strain was named as Pericampylus glaucus witches’-broom (PgWB) phytoplasma, PgWB-hnda strain, belonging to 16SrI-B subgroup by iPhyClassifier analysis. Homology and phylogenetic analysis indicated that based on 16S rRNA gene fragments, PgWB-hnda, pepper yellow crinkle phytoplasma PYC-hnhk (MT760793), chinaberry witches’-broom phytoplasma CWB-hnsy1 (KP662119) and CWB-hn (EF990733), periwinkle virescence phytoplasma PeV-hnhk (KP662136), with 100.0 % identity value, arecanut yellow leaf phytoplasma AYL-hnwn (FJ998269) and AYL-hn (FJ694685), with 99.8 % identity value, were clustered into one clade. Based on the analysis of tuf gene sequence fragments, PgWB was closely related to PYC-hnhk (MT755960), CWB-hnsy1 (KP662155), PeV-hnhk (KP662172) with 99.9 % identity value. Based on the analysis of secA gene sequence fragments, PgWB was closely related to CWB-hnsy1 (KP662173) with 99.7 % identity value, PYC-hnhk (MT755961), PeV-hnhk (KP662190) with 99.4 % identity value. To our knowledge, this is the first time that Pericampylus glaucus witches’-broom disease caused by 16SrI-B subgroup phytoplasma strain was found in China. Multilocus sequence analysis showed that PgWB was closely related to the phytoplasma strains causing pepper yellow crinkle, chinaberry witches’-broom, periwinkle virescence and areca palm yellow leaf diseases, all occurred in Hainan Island of China.


Plant Disease ◽  
2020 ◽  
Author(s):  
Shao-shuai Yu ◽  
Qinghua Tang ◽  
Yuan Wu ◽  
Ming-xing Lin ◽  
Rui-ling Zhao ◽  
...  

Trema tomentosa (Roxb.) Hara belonging to Ulmaceae displayed abnormal symptoms including witches’-broom, internode shortening, leaf chlorosis and leaflet that affected seriously their growth causing financial loss and ecological damage in China. During August through September 2020, these plants with the symptoms were first found and collected in Dingan and Qinghai counties of Hainan province, China. PCR were performed using the primers R16mF2/R16mR1 and secAfor1/secArev3 specific for phytoplasma 16S rRNA and secA gene fragments. The two gene fragments of the DNA extracted from the four disease samples were identical, with length of 1303 bp 16S rRNA and 587 bp secA gene fragments. The phytoplasma strain was named as Trema tomentosa witches’-broom (TtWB) phytoplasma, TtWB-hn strain. Phylogenetic and computer-simulated RFLP analyses based on the nearly full-length 16S rRNA gene sequence indicated that the TtWB phytoplasma strain is more closely related to the 16SrXXXII-A subgroup than to the other subgroups within 16SrXXXII group. It may represent a new subgroup, designed as 16SrXXXII-D subgroup, which is distinct from the other phytoplasma subgroups within the 16SrXXXII group. To our knowledge, this is the first report showing the occurrence of the phytoplasma strain belongs to 16SrXXXII-D subgroup associated with witches’-broom disease in Trema tomentosa in China. Genetic analysis indicated that the TtWB strain was closely related to the phytoplasma strains infecting periwinkle, oil palm, coconut palm in Malyasian, Camptotheca acuminate in Yunnan province of China and Elaeocarpus zollingeri in Japan.


2012 ◽  
Vol 62 (Pt_3) ◽  
pp. 705-709 ◽  
Author(s):  
Rong Jin ◽  
Jing Su ◽  
Hong-Yu Liu ◽  
Yu-Zhen Wei ◽  
Qiu-Ping Li ◽  
...  

A novel alphaproteobacterial strain, designated CPCC 100156T, was isolated from a forest soil sample collected from Hainan Island, South China, and subjected to taxonomic investigation using a polyphasic approach. The pink- to rosy-coloured colonies on TSA and YM agar were smooth and moist. Good growth occurred at 28–32 °C and at pH 7.0–7.5. The respiratory quinone was ubiquinone-9. The polar lipids consisted of phosphatidylcholine (PC), hydroxyphosphatidylethanolamine (OH-PE), phosphatidylglycerol (PG), diphosphatidylglycerol (DPG) and two unidentified aminolipids (AL1, AL2), with a minor amount of ninhydrin-positive phosphoglycolipid. (NPG). The major cellular fatty acids were summed feature 8 (C18 : 1ω7c /C18 : 1ω6c) (49.5 %), summed feature 3 (C16 : 1ω7c/C16 : 1ω6c) (22.5 %), and C16 : 0 (14.0 %). The G+C content of the genomic DNA was 70.3 mol%. The organism showed 16S rRNA gene sequence similarity of 97.37 % with Belnapia moabensis DSM 16746T. Phylogenetic analyses based on 16S rRNA gene sequences showed that the isolate belonged to the family Acetobacteraceae and consistently formed a robust cluster with Belnapia moabensis DSM 16746T in the phylogenetic tree. The DNA–DNA hybridization value between the new isolate and Belnapia moabensis DSM 16746T was 45.6 %. On the basis of the taxonomic evidence, it is proposed that strain CPCC 100156T represents a novel species, for which the name Belnapia rosea sp. nov. is proposed. The type strain is CPCC 100156T ( = DSM 23312T = CGMCC 1.10758T). The description of the genus Belnapia is emended accordingly.


2005 ◽  
Vol 55 (1) ◽  
pp. 303-307 ◽  
Author(s):  
Bernd Schneider ◽  
Ester Torres ◽  
María P. Martín ◽  
Manfred Schröder ◽  
Heinz-Dietmar Behnke ◽  
...  

Pinus silvestris and Pinus halepensis trees grown in Germany and Spain, respectively, showing abnormal shoot branching, dwarfed needles and other symptoms were examined for the presence of plant-pathogenic mollicutes (phytoplasmas). While phytoplasmas could not be detected unambiguously with microscopical methods, PCR amplification using universal phytoplasma primers yielded positive results. Samples collected from symptomatic and non-symptomatic plant parts of both symptomatic Pinus silvestris and Pinus halepensis trees tested positive. Also, surrounding non-symptomatic trees proved to be phytoplasma-infected. Comparisons revealed that the 16S rRNA gene sequences of the phytoplasmas identified in Pinus silvestris and Pinus halepensis were nearly identical. However, the pine phytoplasma is only distantly related to other phytoplasmas. The closest relatives are members of the palm lethal yellowing and rice yellow dwarf groups and ‘Candidatus Phytoplasma castaneae’, which share between 94·5 and 96·6 % 16S rRNA gene sequence similarity. From these data it can be concluded that the phytoplasmas identified in the two Pinus species represent a coherent but discrete taxon; it is proposed that this taxon be distinguished at putative species level under the name ‘Candidatus Phytoplasma pini’.


1994 ◽  
Vol 40 (4) ◽  
pp. 313-318 ◽  
Author(s):  
M. Sajjad Mirza ◽  
Dittmar Hahn ◽  
Svetlana V. Dobritsa ◽  
Antoon D. L. Akkermans

Part of the 16S rRNA gene was amplified directly from uncultured endophyte populations within the root nodules of Datisca cannabina and three strains isolated from nodules of Alnus glutinosa (AgKG′84/4), Coriaria nepalensis (Cn3), and D. cannabina (Dc2). Sequence comparison based on 930 nucleotides indicated that the endophyte of D. cannabina nodules belongs to the genus Frankia and is highly related to the endophyte of C. nepalensis root nodules. The relatedness of the endophytes of C. nepalensis and D. cannabina nodules was also reflected by closely related nifH sequences amplified from the nodules. 16S rRNA sequence analysis of the noninfective strains obtained from both D. cannabina (Dc2) and C. nepalensis (Cn3) nodules also revealed the close relationship of these strains to the genus Frankia.Key words: nitrogen fixation, Frankia, 16S rRNA, nifH.


2007 ◽  
Vol 57 (10) ◽  
pp. 2221-2227 ◽  
Author(s):  
Sören Muurholm ◽  
Sylvie Cousin ◽  
Orsola Päuker ◽  
Evelyne Brambilla ◽  
Erko Stackebrandt

Five isolates that were related phylogenetically to members of the genus Pedobacter were isolated from freshwater of the hard-water creek Westerhöfer Bach, North Germany. The five strains (WB 2.1-25T, WB 2.3-71T, WB 3.3-3T, WB 3.3-22T and WB 2.3-45T) were Gram-negative and chemoheterotrophic, with rod-shaped cells. Most of their metabolic properties matched those given in the description of the genus Pedobacter. Consistent with the genus description, their fatty acids included mainly iso-C15 : 0 and summed feature 3 (C16 : 1 ω7c, iso-C15 : 0 2-OH or both); C16 : 1 ω5c, C16 : 0, iso-C15 : 0 3-OH, C16 : 0 3-OH and iso-C17 : 0 3-OH were present in smaller amounts. The major isoprenoid quinone was menaquinone 7. With one exception, binary similarity values of the almost complete 16S rRNA gene sequences determined among the isolates as well as between the isolates and type strains of Pedobacter species were lower than 98.5 %. The only exception was the close relationship between Pedobacter caeni DSM 16990T and strain WB 2.3-45T (99.2 % similarity). DNA–DNA reassociation values determined for this pair of strains was 29.8 %, indicating that strain WB 2.3-45T represents a unique genospecies. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strains WB 2.1-25T and WB 2.3-71T form a group that is moderately related to P. caeni and strain WB 2.3-45T (98.5 % similarity). Strains WB 3.3-3T and WB 3.3-22T (98.5 % similarity) branched separately from these four organisms. The five phylogenetically isolated strains differed from each other as well as from the type strain of the type species (Pedobacter heparinus DSM 2366T) and some related representatives of the genus in several metabolic reactions and cultural parameters. On the basis of phenotypic and phylogenetic distinctiveness, five novel species are proposed: Pedobacter duraquae sp. nov., with WB 2.1-25T (=DSM 19034T=CIP 109481T) as the type strain; Pedobacter westerhofensis sp. nov., with WB 3.3-22T (=DSM 19036T=CIP 109479T) as the type strain; Pedobacter metabolipauper sp. nov., with WB 2.3-71T (=DSM 19035T=CIP 109480T) as the type strain; Pedobacter hartonius sp. nov., with WB 3.3-3T (=DSM 19033T=CIP 109468T) as the type strain; and Pedobacter steynii sp. nov., with WB 2.3-45T (=DSM 19110T=CIP 109507T) as the type strain.


2018 ◽  
Vol 11 (1) ◽  
pp. 74-81
Author(s):  
Oľga Lukačovičová

Abstract The method of samples processing and subsequent their analysis using Radionuclide X-ray Fluorescence Spectrometry for medical plant samples and drugs has been developed. Technical aspects of the measurement such as mutual geometrical arrangement of tablets (of various size and mass), semiconductor Si/Li detector and the primary source of radiation, 238Pu, are discussed. Technique of Radionuclide X-ray Fluorescence Spectrometry (RXFS) has been applied on samples of medical products and medicines used to treat liver diseases. Medicinal herbs, teas in infusion bags and the drug from this group, were analysed in the solid state (in the form of tablets) or as the leachate, applied to the ion exchanger resin. Chelating Extraction Disks EmporeTM have proven the ability to extract elements from liquid samples under certain conditions and thus to prepare samples for RXFS analysis and evaluation. To ensure correct results, an optimal pH environment has been found. The detection limits of selected elements (Cr, Mn, Fe, Ni, Cu, Zn and Pb) have been calculated for both solid and liquid state samples. Standard addition method and calibration curve parameters have been used to evaluate the concentration of each element in studied samples. Determined concentrations of the examined elements were compared with permissible contaminant limits for the relevant food law in Slovak Republic.


2019 ◽  
Vol 48 (2) ◽  
pp. 279-287
Author(s):  
Dongmei Yang ◽  
Xiying Liu ◽  
Pingling Fan ◽  
Yougen Wu

Traditional usage of wild edible plants reflect the dietary habits as they have potential medicinal functions and the awareness of health care of Li Minority in Hainan Island. To facilitate the use and development of wild edible plants in Hainan Island, information was collected four times by semi-structured interviews in 17 traditional markets of Baoting and Lingshui. Twenty six species belonging to 24 genera under 16 families of vascular plants were recognized as wild edible plants. 84.62% of the total species were annual and perennial herbs. Above ground parts were the most frequently used parts. The most common preparation mode was plain-frying. Heat-clearing was the most common medicinal function. Weather, the size of population and plant diversity in local areas have close relationship with the species richness of wild edible plants traded in traditional markets. Geographical difference and time are not critical for the fluctuation of price.


1999 ◽  
Vol 45 (8) ◽  
pp. 639-645 ◽  
Author(s):  
Shotaro Ando ◽  
Tadashi Yokoyama

To elucidate the phylogenetic relationships between Thai soybean bradyrhizobia and USDA strains of Bradyrhizobium, restriction fragment length polymorphism (RFLP) analysis using the nifDK gene probe and sequencing of the partial 16S rRNA gene were performed. In our previous work, Thai isolates of Bradyrhizobium sp. (Glycine max) were separated clearly from Bradyrhizobium japonicum and Bradyrhizobium elkanii based on the RFLP analysis using the nodDYABC gene probe. RFLP analysis using the nifDK gene probe divided 14 Thai isolates and eight USDA strains of B. japonicum into different groups, respectively, but categorized into the same cluster. All of seven strains within these Thai isolates had the same sequence of the partial 16S rRNA gene, and it was an intermediate sequence between those of B. japonicum USDA 110 and B. elkanii USDA 76T. Furthermore, three USDA strains of B. japonicum, USDA 6 (B. japonicum ATCC 10324T), USDA 115 and USDA 129, had the same partial 16S rRNA gene sequence that seven Thai isolates had. These results suggest that Thai isolates of Bradyrhizobium sp. (Glycine max) are genetically distinct from USDA strains of B. japonicum and B. elkanii, but also indicate a close relationship between Thai isolates and USDA strains of B. japonicum.Key words: Bradyrhizobium, 16S rRNA gene, nif gene, soybean, Thailand.


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