Nondisjunction and unequal spindle organization accompany the drive of <i>Aegilops speltoides</i> B chromosomes

Wu, Dandan; Ruban, Alevtina; Fuchs, Joerg; Macas, Jiří; Novák, Petr; Vaio, Magdalena; Zhou, Yong‐Hong; Houben, Andreas

doi:10.1111/nph.15875

Cited by 30 publications

(42 citation statements)

References 61 publications

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“…Mature ovules were fixed in 1× phosphate-buffered saline (PBS) containing 4% paraformaldehyde (PFA) under vacuum at 4 °C for 10 min followed by a 5 h fixation at 4 °C without vacuum. Fixed cowpea ovules were embedded and sectioned according to (Wu et al 2019). For the analysis of pollen mother cells, immature anthers were fixed with 1× PBS containing 4% PFA under vacuum at 4 °C for 10 min followed by a 30 min fixation at 4 °C without vacuum.…”

Section: Methodsmentioning

confidence: 99%

Unequal contribution of two paralogous centromeric histones to function the cowpea centromere

Ishii

Juranić

Maheshwari

et al. 2020

Preprint

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Short title: Functional characterization of cowpea CENH3sOne-sentence summary: The two paralogous centromeric histones (CENH3) of cowpea contribute unequal to the function of the centromere. AbstractThe legume cowpea (Vigna unguiculata, 2n=2x=22) has significant tolerance to drought and heat stress. Here we analysed and manipulated cowpea centromere-specific histone H3 (CENH3) genes, aiming to establish a centromere-based doubled-haploid method for use in genetic improvement of this dryland crop in future. Cowpea encodes two functional CENH3 variants (CENH3.1 and CENH3.2) and two CENH3 pseudogenes. Phylogenetic analysis suggests that gene duplication of CENH3 occurred independently during the speciation of V. unguiculata and the related V. mungo without a genome duplication event. Both functional cowpea CENH3 variants are transcribed, and the corresponding proteins are intermingled in subdomains of different types of centromere sequences in a tissue-specific manner together with the outer kinetochore protein CENPC. CENH3.2 is removed from the generative cell of mature pollen, while CENH3.1 persists. Differences between both CENH3 paralogs are restricted to the N-terminus. The complete CRISPR/Cas9-based inactivation of CENH3.1 resulted in delayed vegetative growth and sterility, indicating that this variant is needed for plant development and reproduction. By contrast, CENH3.2 knockout individuals did not show obvious defects during vegetative and reproductive development, suggesting that the gene is an early stage of subfunctionalization or pseudogenization.

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Section: Methodsmentioning

confidence: 99%

Unequal contribution of two paralogous centromeric histones to function the cowpea centromere

Ishii

Juranić

Maheshwari

et al. 2020

Preprint

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“…At the beginning of the tillering stage, plants were kept for one month at 12 °C to ensure synchronous flowering. Presence of Bs in plants was identified by PCR with primers for the B-specific repeat AesTR-183 [ 26 ] ( Table S1, Supplementary Materials ) and the exact number of Bs was determined by flow-cytometric analysis according to [ 27 ]. Only plants with 2, 3 or 4 Bs were used for the generation of embryos.…”

Section: Methodsmentioning

confidence: 99%

Tissue-Specific Transcriptome Analysis Reveals Candidate Transcripts Associated with the Process of Programmed B Chromosome Elimination in Aegilops speltoides

Boudichevskaia

Ruban

Thiel

et al. 2020

IJMS

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Some eukaryotes exhibit dramatic genome size differences between cells of different organs, resulting from programmed elimination of chromosomes. Here, we present the first transcriptome analysis of programmed chromosome elimination using laser capture microdissection (LCM)-based isolation of the central meristematic region of Aegilops speltoides embryos where B chromosome (B) elimination occurs. The comparative RNA-seq analysis of meristematic cells of embryos with (Bplus) and without Bs (B0) allowed the identification of 14,578 transcript isoforms (35% out of 41,615 analyzed transcript isoforms) that are differentially expressed during the elimination of Bs. A total of 2908 annotated unigenes were found to be up-regulated in Bplus condition. These genes are either associated with the process of B chromosome elimination or with the presence of B chromosomes themselves. GO enrichment analysis categorized up-regulated transcript isoforms into 27 overrepresented terms related to the biological process, nine terms of the molecular function aspect and three terms of the cellular component category. A total of 2726 annotated unigenes were down-regulated in Bplus condition. Based on strict filtering criteria, 341 B-unique transcript isoforms could be identified in central meristematic cells, of which 70 were functionally annotated. Beside others, genes associated with chromosome segregation, kinetochore function and spindle checkpoint activity were retrieved as promising candidates involved in the process of B chromosome elimination.

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“…In Ae. speltoides , for instance, Bs accumulate in the generative nuclei to more than 93% [Wu et al, 2019]. In order to decipher the molecular biology behind the process of B chromosome drive, Anastassia Boudichevskaia (Gatersleben, Germany) demonstrated how comparative RNAseq analysis was used to identify rye Bdrive-associated transcripts.…”

Section: Segregation Behavior and Drive Of B Chromosomesmentioning

confidence: 99%

Addressing Long-Standing Questions with Advanced Approaches: The 4th B Chromosome Conference

Martins¹,

Trifonov²,

Houben³

2020

Cytogenet Genome Res

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B chromosomes (Bs) are enigmatic accessory genomic elements extensively characterized in diverse eukaryotes. Since their discovery in the beginning of the 20th century, B chromosomes have been the subject of investigation in laboratories all around the world. As a consequence, scientific meetings have dealt with B chromosomes, including the most specific and important conference in the field, “The B Chromosome Conference.” The 4th B Chromosome Conference (4BCC) took place in Botucatu, Brazil, in 2019 and was an excellent opportunity to discuss the latest developments in the B chromosome research field. B chromosome science has advanced from classical and molecular cytogenetics to genomics and bioinformatics approaches. The recent advances in next-generation sequencing technologies and high-throughput molecular biology protocols have led Bs to be the subject of massive data analysis, thus enabling the investigation of structural and functional issues not considered before. Although extensive progress has been made, questions are still remaining to be answered. The advances in functional studies based on RNA, epigenetics, and gene ontologies open the perspective to a better understanding of the complex biology of B chromosomes.

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Nondisjunction and unequal spindle organization accompany the drive of Aegilops speltoides B chromosomes

Cited by 30 publications

References 61 publications

Unequal contribution of two paralogous centromeric histones to function the cowpea centromere

Unequal contribution of two paralogous centromeric histones to function the cowpea centromere

Tissue-Specific Transcriptome Analysis Reveals Candidate Transcripts Associated with the Process of Programmed B Chromosome Elimination in Aegilops speltoides

Addressing Long-Standing Questions with Advanced Approaches: The 4th B Chromosome Conference

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