scholarly journals MutS-Homolog2silencing generates tetraploid meiocytes in tomato (Solanum lycopersicum)

2017 ◽  
Author(s):  
Supriya Sarma ◽  
Arun Kumar Pandey ◽  
Maruthachalam Ravi ◽  
Yellamaraju Sreelakshmi ◽  
Rameshwar Sharma
Keyword(s):  
Mismatch Repair ◽  
Protein Function ◽  
Core Protein ◽  
Male Meiosis ◽  
Root Tips ◽  
C Protein ◽  
Ploidy Stability ◽  
Msh2 Protein ◽  
Pollen Formation ◽  
Mother Cells

SUMMARYMSH2 is the core protein of MutS-homolog family involved in recognition and repair of the errors in the DNA. While other members of MutS-homolog family reportedly regulate mitochondrial stability, meiosis, and fertility, MSH2 is believed to participate mainly in mismatch repair. The search for polymorphism inMSH2sequence in tomato accessions revealed both synonymous and nonsynonymous SNPs; however, SIFT algorithm predicted that none of the SNPs influenced MSH2 protein function. The silencing ofMSH2gene expression by RNAi led to phenotypic abnormalities in highly-silenced lines, particularly in the stamens with highly reduced pollen formation.MSH2silencing exacerbated formation of UV-B induced thymine dimers and blocked light-induced repair of the dimers. TheMSH2silencing also affected the progression of male meiosis to a varying degree with either halt of meiosis at zygotene stage or formation of diploid tetrads. The immunostaining of male meiocytes with centromere localized CENPC (Centromere protein C) protein antibody showed the presence of 48 univalent along with 24 bivalent chromosomes suggesting abnormal tetraploid meiosis. The mitotic cells of root tips of silenced lines showed diploid nuclei but lacked intervening cell plates leading to cells with syncytial nuclei. Thus we speculate that tetraploid pollen mother cells may have arisen due to the fusion of syncytial nuclei before the onset of meiosis. It is likely that in addition to Mismatch repair (MMR), MSH2 may have an additional role in regulating ploidy stability.

scholarly journals A study of deregulated MMR pathways and anticancer potential of curcuma derivatives using computational approach

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Priyanjali Bhattacharya ◽  
Trupti N. Patel
Keyword(s):  
Mismatch Repair ◽  
Protein Function ◽  
Curcuma Longa ◽  
Signaling Cascades ◽  
Altered Expression ◽  
Anticancer Properties ◽  
Medicinal Properties ◽  
Multiple Signaling

AbstractPlant derived products have steadily gained momentum in treatment of cancer over the past decades. Curcuma and its derivatives, in particular, have diverse medicinal properties including anticancer potential with proven safety as supported by numerous in vivo and in vitro studies. A defective Mis-Match Repair (MMR) is implicated in solid tumors but its role in haematologic malignancies is not keenly studied and the current literature suggests that it is limited. Nonetheless, there are multiple pathways interjecting the mismatch repair proteins in haematologic cancers that may have a direct or indirect implication in progression of the disease. Here, through computational analysis, we target proteins that are involved in rewiring of multiple signaling cascades via altered expression in cancer using various curcuma derivatives (Curcuma longa L. and Curcuma caesia Roxb.) which in turn, profoundly controls MMR protein function. These biomolecules were screened to identify their efficacy on selected targets (in blood-related cancers); aberrations of which adversely impacted mismatch repair machinery. The study revealed that of the 536 compounds screened, six of them may have the potential to regulate the expression of identified targets and thus revive the MMR function preventing genomic instability. These results reveal that there may be potential plant derived biomolecules that may have anticancer properties against the tumors driven by deregulated MMR-pathways.

scholarly journals Male meiosis and pollen morphology in diploid Indonesian wild bananas and cultivars

The Nucleus ◽  
2021 ◽  
Author(s):  
Fajarudin Ahmad ◽  
Yuyu S. Poerba ◽  
Gert H. J. Kema ◽  
Hans de Jong
Keyword(s):  
Hybrid Sterility ◽  
Pollen Grains ◽  
Chromosome Analysis ◽  
Male Meiosis ◽  
Enzyme Treatment ◽  
Unreduced Gamete ◽  
Ploidy Levels ◽  
Scientific Disciplines ◽  
Sterile Pollen ◽  
Mother Cells

AbstractBreeding of banana is hampered by its genetic complexity, structural chromosome rearrangements and different ploidy levels. Various scientific disciplines, including cytogenetics, linkage mapping, and bioinformatics, are helpful tools in characterising cultivars and wild relatives used in crossing programs. Chromosome analysis still plays a pivotal role in studying hybrid sterility and structural and numerical variants. In this study, we describe the optimisation of the chromosome spreading protocol of pollen mother cells focusing on the effects of standard fixation methods, duration of the pectolytic enzyme treatment and advantages of fluorescence microscopy of DAPI stained cell spreads. We demonstrate the benefits of this protocol on meiotic features of five wild diploid Musa acuminata bananas and a diploid (AA) cultivar banana “Rejang”, with particular attention on pairing configurations and chromosome transmission that may be indicative for translocations and inversions. Pollen slides demonstrate regular-shaped spores except “Rejang”, which shows fertile pollen grains of different size and sterile pollen grains, suggesting partial sterility and unreduced gamete formation that likely resulted from restitutional meiotic divisions.

CYTOLOGY OF 2n POLLEN FORMATION IN DIPLOID ALFALFA, MEDICAGO SATIVA

1979 ◽  
Vol 21 (4) ◽  
pp. 525-530 ◽  
Author(s):  
Nicholi Vorsa ◽  
E. T. Bingham
Keyword(s):  
Medicago Sativa ◽  
Seed Set ◽  
Pollen Grains ◽  
Genetic Constitution ◽  
2N Pollen ◽  
Diploid Parent ◽  
2N Gamete ◽  
Medicago Sativa L ◽  
Pollen Formation ◽  
Mother Cells

Four diploid (2x) clones of alfalfa, Medicago sativa L., which produced good seed set when used as male parents in 4x-2x crosses were selected for study. The 2x clones descended from 2x haploids of cultivated 4x alfalfa. Fertility in the 4x-2x cross was due to the production of pollen with the unreduced chromosome number (2n pollen) from the 2x parent. The cytological mechanism of 2n pollen formation was found to be disorientation of spindles at metaphase II in up to 38% of the pollen mother cells. Thus, both n and 2n pollen were produced by all four diploids examined. Normal spindles at metaphase II were oriented such that they defined the poles of a tetrahedron and resulted in normal tetrads in a tetrahedral arrangement. Disoriented spindles were basically parallel to each other and resulted in formation of dyads and occasionally a triad. Dyads developed into two 2n pollen grains; triads developed into one 2n and two n pollen grains. Since both n and 2n pollen grains are produced by the diploids, they can be maintained as diploids or they can be used as male parents in crosses to tetraploids. The genetic constitution of 2n pollen resulting from parallel spindles is similar to that expected after first division restitution of meiosis and much of the heterozygosity of the diploid parent is conserved in the gametes. The 2n gamete mechanism has potential application in germplasm transfer and in maximizing heterozygosity in tetraploid hybrids.

scholarly journals When Coupled to Natural Transformation in Acinetobacter sp. Strain ADP1, PCR Mutagenesis Is Made Less Random by Mismatch Repair

2005 ◽  
Vol 71 (11) ◽  
pp. 7610-7612 ◽  
Author(s):  
Alison Buchan ◽  
L. Nicholas Ornston
Keyword(s):  
Structure Function ◽  
Mismatch Repair ◽  
Protein Function ◽  
Natural Transformation ◽  
Function Analysis ◽  
Repair System ◽  
Loss Of Function ◽  
Nucleotide Substitutions ◽  
Full Spectrum ◽  
Mismatch Repair System

ABSTRACT Random PCR mutagenesis is a powerful tool for structure-function analysis of targeted proteins, especially when coupled with DNA integration through natural transformation followed by selection for loss of function. The technique has been applied successfully to structure-function analysis of transcriptional regulators, enzymes, and transporters in Acinetobacter sp. strain ADP1. However, the mismatch repair system prevents the full spectrum of nucleotide substitutions that may be selected at the level of protein function from being recovered. This barrier may be overcome by introducing PCR-mutagenized genes into strains in which the corresponding genes have been deleted.

scholarly journals The N-terminal half of the core protein of hepatitis C virus is sufficient for nucleocapsid formation

2004 ◽  
Vol 85 (4) ◽  
pp. 971-981 ◽  
Author(s):  
Nathalie Majeau ◽  
Valérie Gagné ◽  
Annie Boivin ◽  
Marilène Bolduc ◽  
Josée-Anne Majeau ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Core Protein ◽  
Host Cells ◽  
Yeast Cells ◽  
Main Function ◽  
C Protein ◽  
Multifunctional Protein ◽  
The Core

The core (C) protein of hepatitis C virus (HCV) appears to be a multifunctional protein that is involved in many viral and cellular processes. Although its effects on host cells have been extensively discussed in the literature, little is known about its main function, the assembly and packaging of the viral genome. We have studied the in vitro assembly of several deleted versions of recombinant HCV C protein expressed in E. coli. We demonstrated that the 75 N-terminal residues of the C protein were sufficient to assemble and generate nucleocapsid-like particles (NLPs) in vitro. However, homogeneous particles of regular size and shape were observed only when NLPs were produced from at least the first 79 N-terminal amino acids of the C protein. This small protein unit fused to the endoplasmic reticulum-anchoring domain also generated NLPs in yeast cells. These data suggest that the N-terminal half of the C protein is important for formation of NLPs. Similarities between the HCV C protein and C proteins of other members of the Flaviviridae are discussed.

Mitotic and meiotic irregularities in somatic hybrids of Lycopersicon esculentum and Solanum tuberosum

Genome ◽  
1994 ◽  
Vol 37 (5) ◽  
pp. 726-735 ◽  
Author(s):  
A. M. A. Wolters ◽  
H. C. H. Schoenmakers ◽  
S. Kamstra ◽  
J. van Eden ◽  
M. Koornneef ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
High Frequency ◽  
Somatic Hybrids ◽  
Unreduced Gametes ◽  
Root Tip ◽  
Root Tips ◽  
Tetrad Stage ◽  
Chromosome Arm ◽  
Mother Cells

Chromosome numbers were determined in metaphase complements of root-tip meristems of 107 tomato (+) potato somatic hybrids, obtained from five different combinations of parental genotypes. Of these hybrids 79% were aneuploid, lacking one or two chromosomes in most cases. All four hybrids that were studied at mitotic anaphase of root tips showed laggards and bridges, the three aneuploids in a higher frequency than the single euploid. Hybrid K2H2-1C, which showed the highest percentage of aberrant anaphases, possessed 46 chromosomes. Fluorescence in situ hybridization with total genomic DNA showed that this hybrid contained 23 tomato, 22 potato, and 1 recombinant chromosome consisting of a tomato chromosome arm and a potato chromosome arm. The potato parent of K2H2-1C was aneusomatic in its root tips with a high frequency of monosomic and trisomic cells and a relatively high frequency of cells with one fragment or telosome. Meiotic analyses of three tomato (+) potato somatic hybrids revealed laggards, which occurred most frequently in the triploid hybrids, and bridges, which were frequently present in pollen mother cells (PMCs) at anaphase I of hypotetraploid K2H2-1C. We observed putative trivalents in PMCs at diakinesis and metaphase I of eutriploid A7-82A and quadrivalents in part of the PMCs of hypotetraploid K2H2-1C, suggesting that homoeologous recombination between tomato and potato chromosomes occurred in these hybrids. All three hybrids showed a high percentage of first division restitution, giving rise to unreduced gametes. However, shortly after the tetrad stage all microspores completely degenerated, resulting in exclusively sterile pollen.Key words: tomato, potato, symmetric somatic hybrids, chromosomal irregularities, genomic in situ hybridization.

scholarly journals Analysis of alternative splicing events in the root tips and nodules of Pisum sativum L

2019 ◽  
Vol 17 (1) ◽  
pp. 53-63 ◽  
Author(s):  
Evgeny A. Zorin ◽  
Olga A. Kulaeva ◽  
Alexey M. Afonin ◽  
Vladimir A. Zhukov ◽  
Igor A. Tikhonovich
Keyword(s):  
Alternative Splicing ◽  
Functional Groups ◽  
Protein Function ◽  
Cellular Response ◽  
Target Genes ◽  
Intron Retention ◽  
Post Inoculation ◽  
Root Tips ◽  
Symbiotic Relationships ◽  
The Moment

Background. Legumes establish symbioses with nitrogen-fixing bacteria from the Rhizobium group. In exchange for nutrients, bacteria provide fixed nitrogen needed to support plant growth. At the moment, information about the involvement of alternative splicing (AS) in the establishment and maintenance this symbiotic relationships is almost absent, but, as it is a powerful mechanism for the regulation of proteome diversity of the cell, it therefore may participate in cellular response to microsymbionts. Materials and methods. Alternative splicing was analyzed using the assembly of supertranscripts and alignment of the reads from nodules and root tips to this reference. Target genes expression levels was estimated in tips of non-inoculated roots, and in nodules (2, 4, and 6 weeks post inoculation) with use of RT-qPCR. Results.In this study, the analysis of AS events in the nodules and root tips of the pea was carried out. The presence of isoforms of four pea genes (PsSIP1, PsIGN, PsWRKY40, PsPR-10) was confirmed and their expression level was estimated. Conclusion. Pea nodules were shown to be more enriched with AS events compared to root tips. Among the functional groups of genes that demonstrate AS events, one of the most enriched functional groups is the pathogens stress response. Intron retention probably leads to degradation of the transcript via NMD-system or to change of the protein function, that modulates the activity of genes in nodules.

scholarly journals Cytological Analysis of a Clethra alnifolia `Hokie Pink' × C. pringlei Hybrid

HortScience ◽  
2005 ◽  
Vol 40 (2) ◽  
pp. 339-342
Author(s):  
Sandra M. Reed
Keyword(s):  
Chromosome Number ◽  
Cold Hardiness ◽  
Male Gamete ◽  
Cytological Analysis ◽  
Root Tips ◽  
Chromosome Counts ◽  
Pollen Mother Cells ◽  
Size And Shape ◽  
Mother Cells ◽  
Deciduous Shrub

Clethra alnifolia L., a native deciduous shrub cultivated as an ornamental, was recently hybridized with C. pringlei S. Wats. The purpose of this hybridization was to combine the cold hardiness and adaptability of C. alnifolia with the ornamental foliage of C. pringlei. While most of the C. alnifolia × C. pringlei hybrids more closely resembled C. alnifolia than the paternal species, a `Hokie Pink' × C. pringlei hybrid (NA71586) with foliage that flushes red like C. pringlei was recovered. The objectives of this study were to analyze cytologically the F1 and produce a F2 population from NA71586. Chromosome counts from root tips cells indicated that NA71586 has 32 chromosomes. Since the chromosome number of C. alnifolia is 2n = 32 and that of C. pringlei was found to be 2n = 16, NA71586 appears to have developed following fertilization of a C. alnifolia egg with an unreduced male gamete from C. pringlei. Both `Hokie Pink' and C. pringlei exhibited primarily bivalent pairing in pollen mother cells (PMCs). Over half of the PMCs from NA71586 contained 16 bivalents, indicating substantial homology within the C. alnifolia genome. It was theorized that C. alnifolia is either an autotetraploid that exhibits bivalent pairing or a segmental allotetraploid produced from hybridization of species with similar genomes. More than 700 F2 progeny were obtained from self-pollination of NA71586. Although many of the F2 progeny resembled NA71586, variation in foliage color, size and shape was apparent in the population.

scholarly journals Arabidopsis adaptor protein 1G2 is required for female and male gametogenesis

2019 ◽  
Author(s):  
Yongmei Zhou ◽  
Wenqin Fang ◽  
Li-Yu Chen ◽  
Neha Pandey ◽  
Azam Syed Muhammad ◽  
...  
Keyword(s):  
Male Gametophyte ◽  
Large Subunit ◽  
Adaptor Protein ◽  
Complementation Test ◽  
Female Sterility ◽  
Root Tips ◽  
Male Gametogenesis ◽  
Native Promoter ◽  
Male Gametes ◽  
Mother Cells

Abstract Background: The gametophyte s are essential for the productive process in angiosperms. During sexual reproduction in flowering plants, haploid spores are formed from meioses of spore mother cells. The spores then undergo mitosis and develop into female and male gametes and give rise to seeds after fertilization. Results: We identified a female sterile mutant from EMS mutagenesis, and a BC1F2 population was generated for map based cloning of the causal gene. Genome re-sequencing of mutant and non-mutant pools revealed a candidate gene, AP1G2 . Analyses of two insertions mutants, ap1g2-1 +/- in exon 7 and ap1g2-3 -/- in 3’ UTR, revealed partial female sterility. Complementation test using native promoter of AP1G2 restored the function in ap1g2-1 +/- and ap1g2-3 -/- . AP1G2 is a paralog of AP1G1 , encoding the large subunit (γ) of adaptor protein-1 (AP-1). ap1g2 mutation led to defective female and male gametophyte development was determined. In the ap1g2 mutants, the mitotic cycles and synchronic development of female gametophytes were impaired, which led to the arrest of female gametophytes at one nucleus stage FG1. Pollen development in ap1g2 was also arrested at one nucleus stage before PMI (pollen mitosis I). AP1G2 was expressed at high levels in different stages of ovule and pollens and actively dividing tissues, including shoot apical meristems, leaf primordial and root tips. Conclusions: AP1G2 was identified to have a role in the processes of female and male gametogenesis by regulating the first mitosis at one nucleus stage, and the expression pattern suggested AP1G2 is crucial for plant growth and development. Keywords: Arabidopsis, AP1G2 , megagametogenesis, microgametogenesis, development.

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