Analysis of mitochondrial DNA, chloroplast DNA, and double-stranded RNA in fertile and cytoplasmic male-sterile sunflower (Helianthus annuus)

1986 ◽  
Vol 28 (1) ◽  
pp. 121-129 ◽  
Author(s):  
Gregory G. Brown ◽  
Howard Bussey ◽  
Lee J. DesRosiers
Keyword(s):  
Male Sterility ◽  
Restriction Analysis ◽  
Nuclear Gene ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Male Sterile ◽  
Gene Effects ◽  
Mitochondrial Dnas ◽  
Specific Association ◽  
Chloroplast Dnas

The extent of variation in the mitochondrial DNAs (mtDNAs), chloroplast DNAs (ctDNAs), and double-stranded RNAs (dsRNAs) of sunflower lines carrying fertile and male-sterility conferring cytoplasms was examined. To minimize nuclear gene effects, efforts were concentrated on two chromosomally isogenic lines, CM400 (fertile) and cmsCM400 (male sterile), which differ only in their cytogenes. A circular 1.45 kilobases (kb) plasmid DNA was found in the mitochondria of the four fertile lines examined, but was absent in the male-sterile line. Restriction enzyme analysis of mtDNAs of the fertile and male-sterile cytoplasms with BamHI, EcoRI, and HindIII revealed no fragment mobility differences between them other than those which could be ascribed to the 1.45-kb circle. Similar restriction analysis of ctDNA showed no differences between fertile and male-sterile cytoplasms. Both CM400 and cmsCM400 contain dsRNA molecules. The number and sizes of these dsRNAs varied from preparation to preparation in both lines. Species of 3.3 and 1.5 kb, which were the only dsRNAs common to all preparations from CM400, were also the only species common to all preparations from cmsCM400. Thus, no consistent differences between the fertile and male-sterile cytoplasms were seen in these molecules. The specific association of the 1.45-kb plasmid with fertile cytoplasm together with the absence of variation in ctDNA and dsRNA, suggests the involvement of mtDNA in sunflower cytoplasmic male sterility.Key words: DNA (mitochondrial), sterility (male), sterility (cytoplasmic), Helianthus, sunflower, DNA chloroplast.

Mitochondrial DNA rearrangements and transcriptional alterations in the male-sterile cytoplasm of Ogura radish

1988 ◽  
Vol 8 (4) ◽  
pp. 1474-1480
Author(s):  
C A Makaroff ◽  
J D Palmer
Keyword(s):  
Cytoplasmic Male Sterility ◽  
Male Sterility ◽  
Mitochondrial Genes ◽  
Nuclear Genes ◽  
Male Sterile ◽  
Transcriptional Pattern ◽  
Transcriptional Alterations ◽  
Mitochondrial Dnas ◽  
Male Sterile Cytoplasm ◽  
Chloroplast Dnas

Maternally inherited mutations, such as cytoplasmic male sterility, provide useful systems in which to study the function of plant mitochondrial genomes and also their interaction with nuclear genes. We have studied the organization and expression of the organelle genomes of the male-sterile cytoplasm of Ogura radish and compared them with those of normal radish to identify alterations that might be involved in cytoplasmic male sterility. The chloroplast DNAs of Ogura and normal radish are virtually indistinguishable, whereas their mitochondrial DNAs are highly rearranged. Alignment of a restriction map constructed for the 257-kilobase Ogura mitochondrial genome with that published for the 242-kilobase genome of normal radish reveals that the two mitochondrial DNAs differ in arrangement by at least 10 inversions. The transcriptional patterns of several known mitochondrial genes and of rearranged mitochondrial sequences were examined in three nuclear backgrounds. Altered transcripts were observed for three mitochondrial genes, atpA, atp6, and coxI. Rearrangements map near each of these genes and therefore may be responsible for their transcriptional alterations. Radish nuclear genes that restore fertility to the Ogura cytoplasm have no effect on the atp6 and coxI transcripts, but do influence the atpA transcriptional pattern.

scholarly journals Mitochondrial DNA rearrangements and transcriptional alterations in the male-sterile cytoplasm of Ogura radish.

1988 ◽  
Vol 8 (4) ◽  
pp. 1474-1480 ◽  
Author(s):  
C A Makaroff ◽  
J D Palmer
Keyword(s):  
Cytoplasmic Male Sterility ◽  
Male Sterility ◽  
Mitochondrial Genes ◽  
Nuclear Genes ◽  
Male Sterile ◽  
Transcriptional Pattern ◽  
Transcriptional Alterations ◽  
Mitochondrial Dnas ◽  
Male Sterile Cytoplasm ◽  
Chloroplast Dnas

Maternally inherited mutations, such as cytoplasmic male sterility, provide useful systems in which to study the function of plant mitochondrial genomes and also their interaction with nuclear genes. We have studied the organization and expression of the organelle genomes of the male-sterile cytoplasm of Ogura radish and compared them with those of normal radish to identify alterations that might be involved in cytoplasmic male sterility. The chloroplast DNAs of Ogura and normal radish are virtually indistinguishable, whereas their mitochondrial DNAs are highly rearranged. Alignment of a restriction map constructed for the 257-kilobase Ogura mitochondrial genome with that published for the 242-kilobase genome of normal radish reveals that the two mitochondrial DNAs differ in arrangement by at least 10 inversions. The transcriptional patterns of several known mitochondrial genes and of rearranged mitochondrial sequences were examined in three nuclear backgrounds. Altered transcripts were observed for three mitochondrial genes, atpA, atp6, and coxI. Rearrangements map near each of these genes and therefore may be responsible for their transcriptional alterations. Radish nuclear genes that restore fertility to the Ogura cytoplasm have no effect on the atp6 and coxI transcripts, but do influence the atpA transcriptional pattern.

scholarly journals Specific Detection of Xanthomonas axonopodis pv. dieffenbachiae in Anthurium (Anthurium andreanum) Tissues by Nested PCR

2006 ◽  
Vol 72 (2) ◽  
pp. 1072-1078 ◽  
Author(s):  
Isabelle Robène-Soustrade ◽  
Philippe Laurent ◽  
Lionel Gagnevin ◽  
Emmanuel Jouen ◽  
Olivier Pruvost
Keyword(s):  
Diagnostic Tool ◽  
Nested Pcr ◽  
Restriction Analysis ◽  
Detection Threshold ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Amplification Product ◽  
Xanthomonas Axonopodis ◽  
Sequence Characterized Amplified Region ◽  
Pcr Test

ABSTRACT Efficient control of Xanthomonas axonopodis pv. dieffenbachiae, the causal agent of anthurium bacterial blight, requires a sensitive and reliable diagnostic tool. A nested PCR test was developed from a sequence-characterized amplified region marker identified by randomly amplified polymorphic DNA PCR for the detection of X. axonopodis pv. dieffenbachiae. Serological and pathogenicity tests were performed concurrently with the nested PCR test with a large collection of X. axonopodis pv. dieffenbachiae strains that were isolated worldwide and are pathogenic to anthurium and/or other aroids. The internal primer pair directed amplification of the expected product (785 bp) for all 70 X. axonopodis pv. dieffenbachiae strains pathogenic to anthurium tested and for isolates originating from syngonium and not pathogenic to anthurium. This finding is consistent with previous studies which indicated that there is a high level of relatedness between strains from anthurium and strains from syngonium. Strains originating from the two host genera can be distinguished by restriction analysis of the amplification product. No amplification product was obtained with 98 strains of unrelated phytopathogenic bacteria or saprophytic bacteria from the anthurium phyllosphere, except for a weak signal obtained for one X. axonopodis pv. allii strain. Nevertheless, restriction enzyme analysis permitted the two pathovars to be distinguished. The detection threshold obtained with pure cultures or plant extracts (103 CFU ml−1) allowed detection of the pathogen from symptomless contaminated plants. This test could be a useful diagnostic tool for screening propagation stock plant material and for monitoring international movement of X. axonopodis pv. dieffenbachiae.

scholarly journals Restriction enzyme analysis of the chloroplast DNA of Phaseolus vulgaris L. vr. Rio Negro

1996 ◽  
Vol 26 (3) ◽  
pp. 507-509
Author(s):  
Sergio Echeverrigaray ◽  
Maria Tereza Vitral Carvalho ◽  
Eric Derbyshire
Keyword(s):  
Phaseolus Vulgaris ◽  
Chloroplast Dna ◽  
Sucrose Gradient ◽  
Restriction Analysis ◽  
Gradient Centrifugation ◽  
Repeat Sequence ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Phaseolus Vulgaris L ◽  
Rio Negro

The chloroplast DNA of Phaseolus vulgaris L. vr. Rio Negro was isola ted from chloroplasts obtained by descontiuous sucrose gradient centrifugation. The restriction analysis with the enzymes HindIII, EcoRI and BamHI and their combination, allowed to identified more than 20 fragments of 18 to 0.65kb. The size of Phaseolus vulgaris L. cp DNA was estimated in 140kb with the presence of a repeat sequence of about 22kb.

scholarly journals Characterization of cytomegalovirus isolates from patients with AIDS by DNA restriction analysis

1988 ◽  
Vol 101 (3) ◽  
pp. 483-494 ◽  
Author(s):  
D. L. Taylor ◽  
D. Taylor-Robinson ◽  
D. J. Jeffries ◽  
A. S. Tyms
Keyword(s):  
Restriction Analysis ◽  
Genome Structure ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Migration Patterns ◽  
Disease Pattern ◽  
Dna Restriction ◽  
Acquired Immunodeficiency ◽  
Different Strains ◽  
Immunodeficiency Syndrome

SUMMARYThirty-seven isolates of cytomegalovirus (CMV) were obtained from a group of 20 promiscuous homosexual men, either suffering from the acquired immunodeficiency syndrome (AIDS) at the time of CMV isolation, or who developed AIDS subsequently. The isolates of CMV were characterized by the method of DNA restriction analysis. All epidemiologically unrelated strains of CMV exhibited different fragment migration patterns and no one strain appeared to be associated with AIDS or any particular disease pattern in these patients.Sequential isolates of CMV were obtained from nine patients in the study group either from different sites at the same time or from the same site on different dates. In the case of seven of the men, viruses with minor differences in restriction profile were obtained, possiblyrepresenting sub-populations of an endogenous strain of CMV. In two of the patients, reinfection with different strains was apparent. We conclude that reinfections with CMV in AIDS patients can occur, but the isolation of strains exhibiting major differences in genome structure seen by restriction enzyme analysis was uncommon.

scholarly journals Inheritance of Male Sterility in Lesquerella fendleri

HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 483C-483
Author(s):  
D.A. Dierig ◽  
P.M. Tomasi ◽  
T.A. Coffelt
Keyword(s):  
Fatty Acids ◽  
Male Sterility ◽  
Seed Oil ◽  
Nuclear Gene ◽  
Oilseed Crop ◽  
Male Sterile ◽  
Chi Square ◽  
Lesquerella Fendleri ◽  
Alternative Crop ◽  
Hybrid Plants

Lesquerella fendleri (Gray) Wats., Brassicaceae, is a potential oilseed crop native to the southwestern U.S. The seed oil contains hydroxy fatty acids, similar to castor. Unique properties of the oil, along with coproducts, allow additional applications that would not be in competition with castor. Plants with vestigial anthers were discovered in a bulk population growing in the greenhouse in 1993. The inheritance of the trait was investigated the following three crop seasons. Crosses were made among sterile and fertile plants and reciprocals among fertile plants. Chi-square results indicate the male sterility trait is expressed by a recessive nuclear gene with cytoplasmic influence restoring fertility. Cytoplasmic male sterile lines can be utilized for development of hybrids. Development of lines without male sterility should lead to higher yields than current bulk populations of lesquerella. Hybrid plants and higher yields will enhance the commercialization potential of this new, alternative crop.

scholarly journals A simple method for detection of mutations in amino acid 452 of the Spike protein of SARS-CoV-2 using restriction enzyme analysis

2021 ◽  
Vol 62 (4) ◽  
pp. 371-377
Author(s):  
Rossana C. Jaspe ◽  
Yoneira Sulbaran ◽  
Mariana Hidalgo ◽  
Mariana Hidalgo ◽  
Carmen L. Loureiro ◽  
...  
Keyword(s):  
Immune Response ◽  
Amino Acid ◽  
Restriction Enzyme ◽  
Restriction Analysis ◽  
Spike Protein ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Simple Method ◽  
Spike Gene ◽  
Detection Of Mutations

Variants of Concern or Interest of SARS-CoV-2 (VOC or VOI), the coronavirus responsible for COVID-19, have emerged in several countries. Mutations in the amino acid 452 of the Spike protein are particularly important and associated with some of these variants: L452R, present in Delta VOC, and L452Q, present in Lambda VOI. These mutations have been associated with both increased infectivity and evasion of protective immune response. A search on GISAID to detect the number of sequences harboring the L452R mutation and the frequency of Delta VOC among them, showed that since August 2021, most of these sequences belong to the Delta VOC. Restriction enzyme analysis is proposed as a rapid method to detect L452R. A small amplicon from the Spike gene was digested with MspI. A 100% concordance was observed between digestion and sequencing results. The mutation L452Q can also be detected by restriction analysis, allowing the identification of putative Lambda VOIs. The proposed methodology, which allows screening of a great number of samples, could provide a faster information on the prevalence of Delta VOC cases.

scholarly journals Further characterization of 41 isolates of adenovirus types 19/37 by serum neutralization and DNA restriction enzyme analysis

1986 ◽  
Vol 97 (2) ◽  
pp. 377-383 ◽  
Author(s):  
Zong-Da Meng ◽  
Margery L. Kennett ◽  
Suzanne M. Rodger ◽  
Kaye E. Dickson ◽  
Bruce N. Anderson ◽  
...  
Keyword(s):  
Restriction Analysis ◽  
Haemagglutination Inhibition ◽  
Adenovirus Type ◽  
Cross Reactivity ◽  
High Titre ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Serum Neutralization ◽  
Dna Restriction

SummaryForty-one strains of adenovirus type 19/37 (Ad19/37) mainly isolatedI from patients with keratoconjunetivitis or conjunctive between 1974 and 1984 were re-evaluated by serum neutralization (SN), haemagglutination inhibition (HI) and DNA restriction analysis. Of 19 isolates which were neutralized to high titre by antiserum prepared against prototype Ad19, 5 showed cross-reactivity with 32–64 units of Ad37 antiserum, while of 22 strains neutralizedto high titre by Ad37 antiserum, 3 showed eross-reaetivity with 32 units of Ad19 antiserum By DNA restriction analysis, all Ad19 isolates were identieal to each other and to Ad19A virus. Using endonuclease Bgl 1, three variants were observed among the Ad37 isolates.

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