Transplantation and subsequent behavior of mitochondria in cells of Phytophthora

2000 ◽  
Vol 46 (11) ◽  
pp. 992-997 ◽  
Author(s):  
Y H Gu ◽  
W H Ko
Keyword(s):  
Antibiotic Resistance ◽  
Asexual Reproduction ◽  
Second Generation ◽  
Random Distribution ◽  
Phytophthora Capsici ◽  
Selective Medium ◽  
Phytophthora Parasitica ◽  
Subsequent Behavior ◽  
Hybrid Antibiotic ◽  
Fusion Products

Mitochondria isolated from streptomycin-resistant (Sr) protoplasts of Phytophthora parasitica were transferred into chloramphenicol-resistant (Cpr) protoplasts of P. parasitica or Phytophthora capsici with an average successful rate of 1.7 × 10-4, using a selective medium containing streptomycin. No colonies appeared when self-fusion products of donor mitochondria or recipient protoplasts were exposed to the selective medium. Mitochondria isolated from Cpr protoplasts of P. capsici were also transferred into Sr protoplasts of P. parasitica with a similar success rate using a selective medium containing chloramphenicol. Zoospores produced by the Cpr+Sr intraspecific mitochondrial hybrid gave rise to Sr and Cpr+Sr cultures. The second generation zoospores produced by Sr and Cpr+Sr cultures also gave rise to Sr and Cpr+Sr cultures, suggesting the possible occurrence of fusion between some of the Cpr mitochondria and Sr mitochondria, and the displacement of non-fused Cpr mitochondria in the receptor protoplast by the donor Sr mitochondria. Zoospores produced by the interspecific mitochondrial hybrid gave rise to Cpr, Sr, Cpr+Sr, and Cps +Ss cultures. The second generation zoospores produced by Cpr+Sr or Sr cultures also gave rise to the same four types of cultures, suggesting the existence of residual antibiotic-sensitive mitochondria (Cps+Ss) in the parental isolates and the random distribution of Cpr, Sr, and Cps+Ss mitochondria during asexual reproduction. Results suggest that the phenotype of antibiotic resistance / sensitivity was the end result of the interactions among the three types of mitochondria.Key words: mitochondrial transplantation, mitochondrial hybrid, antibiotic resistance, Phytophthora parasitica, Phytophthora capsici.

Segregation following interspecific transfer of isolated nuclei between Phytophthora parasitica and P. capsici

2000 ◽  
Vol 46 (5) ◽  
pp. 410-416 ◽  
Author(s):  
Y H Gu ◽  
W H Ko
Keyword(s):  
Nuclear Transfer ◽  
Phytophthora Capsici ◽  
Selective Medium ◽  
Crossing Over ◽  
Phytophthora Parasitica ◽  
Parasexual Cycle ◽  
Different Types ◽  
Average Success Rate ◽  
Fusion Products ◽  
Interspecific Transfer

Nuclei isolated from metalaxyl-resistant (MR) protoplasts of Phytophthora parasitica were transferred into chloroneb-resistant (CnR) protoplasts of Phytophthora capsici and vice versa, with an average success rate of 2.6 × 10-4 (protoplasts with donor nuclei/regenerated protoplasts), using a selective medium containing only the fungicide tolerated by the nuclear donor. No colonies appeared when self-fusion products of donor nuclei or recipient protoplasts were exposed to the selective medium. Colonies produced by the nuclear transfer formed sectors commonly, and differed from the parental types in appearance. All the zoospores produced by the nuclear hybrids were of normal size, and one-fifth of them contained both MR and CnR genes. Since zoospores are mostly uninucleate, these results indicated the occurrence of chromosome re-assortment or mitotic crossing-over following the production of transitory tetraploids, followed by diploidization during zoosporogenesis, thus suggesting the completion of events leading to a parasexual cycle. Hyphal fragment cultures from a nuclear hybrid tested showed considerable variation in growth rate, mycelial morphology, and level of resistance to metalaxyl, indicating uneven distribution and continuous segregation of different types of nuclei in mycelia during vegetative growth.Key words: interspecific nuclear transfer, parasexual cycle, karyogamy, Phytophthora parasitica,Phytophthora capsici.

Evidence for mitochondrial gene control of mating types in Phytophthora

2005 ◽  
Vol 51 (11) ◽  
pp. 934-940 ◽  
Author(s):  
Yu-Huan Gu ◽  
Wen-Hsiung Ko
Keyword(s):  
Mating Type ◽  
Mitochondrial Gene ◽  
Phytophthora Capsici ◽  
Nuclear Genes ◽  
Gene Control ◽  
Mating Types ◽  
Phytophthora Parasitica ◽  
Decisive Effect ◽  
Mating Type Genes ◽  
Fusion Products

When protoplasts carrying metalaxyl-resistant (Mr) nuclei from the A1 isolate of Phytophthora parasitica were fused with protoplasts carrying chloroneb-resistant (Cnr) nuclei from the A2 isolate of the same species, fusion products carrying Mr nuclei were either the A2 or A1A2 type, while those carrying Cnr nuclei were the A1, A2, or A1A2 type. Fusion products carrying Mr and Cnr nuclei also behaved as the A1, A2, or A1A2 type. The result refutes the hypothesis that mating types in Phytophthora are controlled by nuclear genes. When nuclei from the A1 isolate of P. parasitica were fused with protoplasts from the A2 isolate of the same species and vice versa, all of the nuclear hybrids expressed the mating type characteristics of the protoplast parent. The same was true when the nuclei from the A1 isolate of P. parasitica were fused with the protoplasts from the A0 isolate of Phytophthora capsici and vice versa. These results confirm the observation that mating type genes are not located in the nuclei and suggest the presence of mating type genes in the cytoplasms of the recipient protoplasts. When mitochondria from the A1 isolate of P. parasitica were fused with protoplasts from the A2 isolate of the same species, the mating type of three out of five regenerated protoplasts was changed to the A1 type. The result demonstrated the decisive effect of mitochondrial donor sexuality on mating type characteristics of mitochondrial hybrids and suggested the presence of mating type genes in mitochondria. All of the mitochondrial hybrids resulting from the transfer of mitochondria from the A0 isolate of P. capsici into protoplasts from the A1 isolate of P. parasitica were all of the A0 type. The result supports the hypothesis of the presence of mating type genes in mitochondria in Phytophthora.Key words: mating type, mitochondrial gene, Phytophthora parasitica, Phytophthora capsici.

Creation of hybrid vigor through nuclear transplantation in Phytophthora

2001 ◽  
Vol 47 (7) ◽  
pp. 662-666 ◽  
Author(s):  
Yu-Huan Gu ◽  
Wen-Hsiung Ko
Keyword(s):  
Protoplast Fusion ◽  
Fungicide Resistance ◽  
Phytophthora Capsici ◽  
Hybrid Vigor ◽  
Nuclear Transplantation ◽  
Phytophthora Parasitica ◽  
Isolated Nuclei ◽  
Donor Cells ◽  
Potential Applications ◽  
Fusion Products

When isolated nuclei of a diploid oomycete, Phytophthora parasitica, were fused with protoplasts of another strain of the same species, the regenerated nuclear hybrids grew faster than the parental isolates. Such a phenomenon did not occur in hybrids regenerated from mitochondrion–protoplast or protoplast–protoplast fusion products between these two strains. These results indicate that hybrid vigor is the result of the interaction between two different kinds of nuclei, but not between mitochondria, and they suggest that the presence of mitochondria from nuclear donor cells represses the expression of increased vigor. The nuclear hybrids also expressed increased fungicide resistance and propagule production. Increased vigor in growth was also observed in the interspecific nuclear hybrids when isolated nuclei of P. parasitica were transferred into protoplasts of Phytophthora capsici, and vice versa. This phenomenon may have potential applications, such as the creation of superior fungal strains and plant cultivars with improved commercial traits for usage in industry and agriculture.Key words: hybrid vigor, nuclear transplantation, Phytophthora parasitica, Phytophthora capsici.

scholarly journals Phenotypic assortment in Tetrahymena thermophila: assortment kinetics of antibiotic-resistance markers, tsA, death, and the highly amplified rDNA locus.

Genetics ◽  
1988 ◽  
Vol 120 (2) ◽  
pp. 389-395
Author(s):  
E V Merriam ◽  
P J Bruns
Keyword(s):  
Antibiotic Resistance ◽  
Random Distribution ◽  
Tetrahymena Thermophila ◽  
Single Gene ◽  
Rdna Locus ◽  
Phenotypic Assortment ◽  
Resistance Markers ◽  
Kinetics Of ◽  
Input Ratio ◽  
As If

Abstract Phenotypic assortment in Tetrahymena thermophila results from random distribution of alleles during amitotic division of the macronucleus. The rate of assortment is dependent on input ratio and the number of assorting units. The assortment of the antibiotic resistance markers Chx, Mpr and gal was determined and is consistent for each with the model of 45 assorting chromosomes. The gene tsA (previously ts-1) shows normal assortment, in contrast to previous reports. A mutation in the highly amplified ribosomal locus (rdnA2) assorts as if present at only 45 copies. Death of clones occurred at a rate consistent with assortment for a single gene.

Antibiotic Resistance of Coagulase-Negative Staphylococci and Lactic Acid Bacteria Isolated from Naturally Fermented Chinese Cured Beef

2018 ◽  
Vol 81 (12) ◽  
pp. 2054-2063 ◽  
Author(s):  
JING WANG ◽  
MINGYUE LI ◽  
JING WANG ◽  
MIAOMIAO LIU ◽  
KUN YANG ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Random Amplified Polymorphic Dna ◽  
Selective Medium ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Rrna Gene ◽  
M Gene ◽  
Coagulase Negative Staphylococci

ABSTRACT This study provided phenotypic and molecular analysis of the antibiotic resistance within coagulase-negative staphylococci and lactic acid bacteria isolated from naturally fermented Chinese cured beef. A total of 49 strains were isolated by selective medium and identified at the species level by 16S rRNA gene sequencing as follows: Staphylococcus carnosus (37), Lactobacillus plantarum (6), Weissella confusa (4), Lactobacillus sakei (1), and Weissella cibaria (1). All strains were typed by random amplified polymorphic DNA fingerprinting, and their antibiotic resistances profiles to 15 antibiotics were determined as the MIC by using the agar dilution method. All the tested strains were sensitive to ampicillin, and most of them were also sensitive to penicillin, gentamycin, neomycin, norfloxacin, and ciprofloxacin with low MICs. High resistance to streptomycin, vancomycin, erythromycin, roxithromycin, lincomycin, and kanamycin was widely observed, while the resistant levels to tetracycline, oxytetracycline, and chloramphenicol varied. The presence of corresponding resistance genes in resistant isolates was investigated by PCR, with the following genes detected: tet(M) gene in 9 S. carnosus strains and 1 W. confusa strain; erm(F) gene in 10 S. carnosus strains; ere(A) gene in 6 S. carnosus strains; ere(A) gene in 4 S. carnosus strains and 1 L. plantarum strain; and str(A) gene and str(B) gene in 3 S. carnosus strains. The results indicated that multiple antibiotic resistances were common in coagulase-negative staphylococci and lactic acid bacteria strains isolated from naturally fermented Chinese cured beef. Safety analysis and risk assessment should be performed for application in meat products.

Antibiotic Resistance Profiling of Salmonella sp. isolated from African Catfish (Clarias gariepinus)

2021 ◽  
Vol 16 (3) ◽  
pp. 44-47
Author(s):  
P.T. Fowoyo
Keyword(s):  
Antibiotic Resistance ◽  
Pathogenic Bacteria ◽  
Clarias Gariepinus ◽  
Fish Farming ◽  
Selective Medium ◽  
Diffusion Method ◽  
African Catfish ◽  
Fresh Water Fish ◽  
Peptone Water ◽  
Amoxicillin Clavulanic Acid

The African catfish (Clarias gariepinus) is an important fresh water fish consumed by a large percentage of the populace globally and it may be contaminated by pathogenic bacteria such as Salmonella sp. In this study, a total of fifteen (15) samples of African catfish were collected from different markets in Lokoja, Nigeria. The Salmonella sp. were isolated from the catfish samples by pre-enrichment in peptone water and subsequent inoculation on selective medium namely brilliant-green agar (BGA), bismuth sulphite agar (BSA) and Salmonella-Shigella agar (SSA). The Salmonella isolates were tested for susceptibility to 10 different commercially available antibiotics using the disc diffusion method. A total of thirty-four Salmonella species was isolated. The percentage occurrence of Salmonella sp. in the catfishes examined was very high (80%). The incidence of Salmonella sp. in the intestine (86.7%) of the catfish was higher than for the gills (66.7%) and the skin (73.3%). Majority of the isolates were resistant to Amoxicillin, Sulfomethoxazole-trimethoprim, Amoxicillin-clavulanic acid and Streptomycin. This study therefore demonstrated the occurrence of Salmonella species in African catfish with some exhibiting antibiotic resistance. Thus, there is a potential risk of transmission of drug resistant Salmonella species to man when contaminated catfish is consumed. The use of antibiotics in fish farming should be regulated so as to decrease antibiotic residues in fish.

scholarly journals Comparative study of strains of lactic acid bacteria contained in"Lactos" with imported probiotics

2021 ◽  
Vol 32 (1) ◽  
pp. 12-20
Author(s):  
Demberel Shirchin ◽  
Ulzii-Uchral Davaasuren ◽  
Tsogtbaatar Lkhagvajav ◽  
Bulgan Boldbaatar
Keyword(s):  
Antibiotic Resistance ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Biological Properties ◽  
Selective Medium ◽  
Lactobacillus Helveticus ◽  
Nutrient Media ◽  
Local Strains ◽  
Cultural Properties ◽  
Mrs Medium

We studied some biological properties of the local strains Lactobacillus helveticus LBMA44c and Lactobacillus delbruesckii subsp.bulgaricus LBMA65b contained in the domestic probiotic LACTOSA in compared to imported probiotics Lactobacillus, Linex-malysh and Acidophilus Probiotic Blend.  The strains cultivated in liquid and solid selective MRS medium, obtained as a result of subsequent dilutions of a sample of the probiotics under study, perfectly demonstrated the general features of the morphological and cultural properties of lactic acid bacteria. The authors found that the indicators CFU/g of  LACTOS (1.9-2.0x108 CFU/g) is not less than that of imported ones, in terms of the oxidation of nutrient media, it is the same as that of Acidophilus Probiotic Blend-and slightly lower than that of Linex and Lacto-Fit. Strains of lactic acid bacteria in all studied probiotics are well cultivated at 37, 45, 60 degrees (0C) and on a selective medium with an addition at the rate of 2, 4, 6 percent of natural bile and grows poorly on a medium with the addition of 4 and 6 percent NaCl. We also noted that the local strains in our LACTOS probiotic were relatively more resistant to antibiotics than imported ones, according to antibiotic resistance estimates. Thus, the local strains of lactic acid bacteria contained in the domestic probiotic under the LACTOS trademark do not differ from the imported ones. “Лактос” пробиотикт агуулагдах сүүн хүчлийн бактерийн шинж чанарыг импортынхтой харьцуулсан судалгаа “Лактос” пробиотикт агуулагдах сүүн хүчлийн бактерийн Lаctobacillus helveticus LBMA44c болон Lаctobacillus delbruesckii subsp.bulgaricus LВMА65b нутгийн омгийн эмнэлэг-биологийн чадавхын зарим үзүүлэлтийг импортын Lасто-Fit, Линекс Малыш №20, Acidophilus Probiotic Blend-ийнхтэй харьцуулан судлав. Судалгаанд хамрагдсан пробиотикуудын дээжний шингэлэлтээс MRS -ийн хатуу, шингэн сонгомол орчинд ургуулахад тэдгээр нь сүүн хүчлийн бактерийн өсгөвөржилт, хэлбэр зүйн нийтлэг шинж байдлыг үзүүлжээ. Пробиотикт агуулагдах амилах чадвартай бактерийн эсийн тоогоор Лактосынх (1,9-2,0х108КҮН/г) импортынхоос дутахгүй, харин орчныг хүчилсэг болгох идэвхээрээ Линекс Малыш, Lасtо-Fit-ээс бага зэрэг доогуур Acidophilus Probiotic Blend -тэй ойролцоо байна. Тэдгээр нь Цельсийн 37, 45, 60 хэмийн дулаанд болон 2, 4, 6 хувиар цөс нэмсэн тэжээлт орчинд сайн өсгөвөржих боловч цэвэр NaCl-ийн нэмэлт 4-6 хувь хүрэхэд пробиотикийн агууламж дахь сүүн хүчлийн бактерийн ургалтын эрчим дундаас доош гэсэн үнэлэмж рүү шилжсэн юм. Лактост агуулагдах сүү хүчлийн бактерийн омгууд нь судалгаанд ашигласан 5 төрлийн антибиотик тэсвэрлэх чадавхын дундаж үзүүлэлтээр импортын пробиотикийнхтой ойролцоо ажээ.  Түлхүүр үг: пробиотик, сүүн хүчлийн бактери, тэсвэрт чанар, өсгөвөржилт

scholarly journals EFFECT OF RADIATION DOSAGE ON EFFICIENCY OF CHLOROPLAST TRANSFER BY PROTOPLAST FUSION IN NICOTIANA

Genetics ◽  
1982 ◽  
Vol 100 (3) ◽  
pp. 487-495
Author(s):  
László Menczel ◽  
Gábor Galiba ◽  
Ferenc Nagy ◽  
Pál Maliga
Keyword(s):  
Protoplast Fusion ◽  
Radiation Dosage ◽  
Nicotiana Plumbaginifolia ◽  
Selective Medium ◽  
60Co Source ◽  
Fragmentation Patterns ◽  
Fusion Products ◽  
Lethal Doses ◽  
Nicotiana Tabacum Sr1 ◽  
Ecori Digestion

ABSTRACT Chloroplasts of Nicotiana tabacum SR1 were transferred into Nicotiana plumbaginifolia by protoplast fusion. The protoplasts of the organelle donor were irradiated with different lethal doses using a 60Co source, to facilitate the elimination of their nuclei from the fusion products. After fusion induction, clones derived from fusion products and containing streptomycin-resistant N. tabacum SR1 chloroplasts were selected by their ability to green on a selective medium. When N. tabacum protoplasts were inactivated by iodoacetate instead of irradiation, the proportion of N. plumbaginifolia nuclear segregant clones was low (1-2%). Irradiation markedly increased this value: Using 50, 120, 210 and 300 J kg-1 doses, the frequency of segregant clones was 44, 57, 84 and 70 percent, respectively. Regeneration of resistant N. plumbaginifolia plants with SR1 chloroplasts indicated that plastids can be rescued from the irradiated cells by fusion with untreated protoplasts. Resistant N. plumbaginifolia plants that were regenerated (43 clones studied) had diploid (2n = 2X = 20) or tetraploid chromosome numbers and were identical morphologically to parental plants. The absence of aneuploids suggests that in these clones irradiation resulted in complete elimination of the irradiated N. tabacum nuclei. Resistance is inherited maternally (five clones tested). The demonstration of chloroplast transfer and the presence of N. tabacum plastids in the N. plumbaginifolia plants was confirmed by chloroplast DNA fragmentation patterns after EcoRI digestion.

scholarly journals Buckeye Rot of Tomato Caused by Phytophthora capsici in Michoacan, Mexico

Plant Disease ◽  
2003 ◽  
Vol 87 (7) ◽  
pp. 872-872 ◽  
Author(s):  
S. P. Fernández-Pavía ◽  
G. Rodríguez-Alvarado ◽  
J. M. Sánchez-Yañez
Keyword(s):  
Phytophthora Capsici ◽  
Selective Medium ◽  
The State ◽  
American Phytopathological Society ◽  
Tomato Fruits ◽  
Moist Chamber ◽  
Tomato Production ◽  
Tomato Plants ◽  
Initial Symptoms ◽  
Diseased Fruit

The state of Michoacan is one of the main fresh pepper (Capsicum annuum L.) and tomato (Lycopersicon esculentum Mill.) producers in Mexico. During the last 5 years, pepper-producing areas in the state have become unproductive due to root-rotting pathogens, mainly Phytophthora capsici Leonian. Growers trying to overcome losses have increased tomato production in areas previously used for pepper production. Field-grown tomato plants with diseased green tomato fruits were observed in Tacambaro, Michoacan, during August 2002. Initially, brown-to-black lesions developed on fruits in contact with soil, followed by infection of the upper fruits in the raceme. Lesions enlarged and dark zonate “buckeye” bands were formed in the affected area. Diseased fruit turned mushy. Symptoms observed were similar to those described for buckeye rot of tomato (1). Diseased fruit were surface disinfested with 70% ethanol, cut into 0.5-cm slices, and incubated in a moist chamber to induce mycelial colonization. Isolation from mycelial tufts growing through the tomato slice was performed 3 days later, and mycelia was transferred to PARP selective medium (corn meal agar (CMA) plus ampicillin, pimaricin, rifampicin, and pentachloronitrobenzene). P. capsici was consistently isolated from diseased tomato fruits. Oomycete identification was based on sporangial and gametangial characteristics of cultures grown on CMA (1). Sporangia microscopically observed were spherical, broadly ellipsoid or obovoid with one papilla (occasionally two papillae), and deciduous with a long pedicel. Chlamydospores were not present (2). The isolates were heterothallic, and oogonia with amphigynous antheridia were observed in pairings with A1 and A2 isolates of P. capsici. Three isolates were A1 and two isolates were A2. To confirm pathogenicity, two experiments were performed using 20 healthy unwounded green tomatoes. One isolate of each mating type was tested. Isolates were grown for 5 days on CMA, and fruits were inoculated by placing P. capsici in contact with the fruit. Inoculated fruits were kept in a moist chamber at room temperature (17 to 20°C). Initial symptoms in the form of brown-to-black lesions appeared 24 h after inoculation. One week after inoculation, symptoms were similar to those observed in field-grown plants, and P. capsici was recovered from the margins of the diseased tissue. All inoculated fruits rotted. To our knowledge, this is the first report of P. capsici causing buckeye rot on tomato in Michoacan and of the presence of both mating types in the area. Reference: (1) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society. St. Paul MN, 1996. (2) M. Aragaki and J. Y. Uchida. Mycologia 93:137, 2001.

scholarly journals Phylogenetic Analysis and Antibiotics Resistance of Listeria Monocytogenes Contaminating Chicken Meat in Surabaya, Indonesia

2020 ◽  
Vol 2020 ◽  
pp. 1-7
Author(s):  
Eduardus Bimo Aksono ◽  
Katty Hendriana Priscilla Riwu ◽  
A. T. Soelih Estoepangestie ◽  
Herinda Pertiwi
Keyword(s):  
Phylogenetic Analysis ◽  
Antibiotic Resistance ◽  
Listeria Monocytogenes ◽  
Selective Medium ◽  
Chicken Meat ◽  
Antibiotics Resistance ◽  
Isolation And Identification ◽  
Traditional Markets ◽  
Meat Samples ◽  
New Generation

The objective of this study was to identify the phylogenetic analysis and antibiotic resistance of Listeria monocytogenes contaminating chicken meat in Surabaya. 60 chicken meat samples were collected from supermarkets, mobile vendors, and traditional markets in Surabaya. A selective medium is used for isolation and identification of Listeria monocytogenes by chopping 25 grams of the chicken meat and to put it into the sterilized Erlenmeyer flasks. Some methods were used for the identification procedures, such as biochemical and morphological tests, antibiotic resistance test, PCR, and sequencing; also a phylogenetic analysis was conducted by a neighbor-joining analysis using Genetix Mac ver 8.0 with hlyA genes of Listeria monocytogenes recorded in GenBank, such as Lineage I (KC808543), Lineage II (AY229462, AY229346, AY229499, and AY229404), Lineage III (KJ504139, HQ686043, KJ504116, and DQ988349), and Lineage IV (EU840690, EF030606). The result shows that the prevalence of L. monocytogenes in Surabaya contaminating the chicken meat samples from the supermarkets was 10% (2/20), from the mobile vendors was 0/20 (0%), and from the traditional markets was 5% (1/20). It was seen from the band at 456 bp fragment. Furthermore, three isolates found in Surabaya were included in the new lineages which were resistant to old-generation antibiotics such as sulfamethonazole-trimetophrim (SXT) and amoxyllin sulbactam (MAS), but they were still sensitive to new-generation antibiotics such as cefotaxime (CTX) and meropenem (MEM).

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