PMID- 29760782
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20220317
IS  - 1755-8166 (Print)
IS  - 1755-8166 (Electronic)
IS  - 1755-8166 (Linking)
VI  - 11
DP  - 2018
TI  - An improved method for inducing prometaphase chromosomes in plants.
PG  - 32
LID - 10.1186/s13039-018-0380-6 [doi]
LID - 32
AB  - BACKGROUND: Detailed karyotyping using metaphase chromosomes in melon (Cucumis 
      melo L.) remains a challenge because of their small chromosome sizes and poor 
      stainability. Prometaphase chromosomes, which are two times longer and loosely 
      condensed, provide a significantly better resolution for fluorescence in situ 
      hybridization (FISH) than metaphase chromosomes. However, suitable method for 
      acquiring prometaphase chromosomes in melon have been poorly investigated. 
      RESULTS: In this study, a modified Carnoy's solution II (MC II) [6:3:1 (v/v) 
      ethanol: acetic acid: chloroform] was used as a pretreatment solution to obtain 
      prometaphase chromosomes. We demonstrated that the prometaphase chromosomes 
      obtained using the MC II method are excellent for karyotyping and FISH analysis. 
      We also observed that a combination of MC II and the modified air dry (ADI) 
      method provides a satisfactory meiotic pachytene chromosome preparation with 
      reduced cytoplasmic background and clear chromatin spreads. Moreover, we 
      demonstrated that pachytene and prometaphase chromosomes of melon and Abelia x 
      grandiflora generate significantly better FISH images when prepared using the 
      method described. We confirmed, for the first time, that Abelia x grandiflora has 
      pairs of both strong and weak 45S ribosomal DNA signals on the short arms of 
      their metaphase chromosomes. CONCLUSION: The MC II and ADI method are simple and 
      effective for acquiring prometaphase and pachytene chromosomes with reduced 
      cytoplasm background in plants. Our methods provide high-resolution FISH images 
      that can help accelerate molecular cytogenetic research in plants.
FAU - Setiawan, Agus Budi
AU  - Setiawan AB
AD  - 1Laboratory of Genetics and Plant Breeding, Graduate School of Horticulture, 
      Chiba University, Matsudo, Chiba 271-8510 Japan. ISNI: 0000 0004 0370 1101. GRID: 
      grid.136304.3
FAU - Teo, Chee How
AU  - Teo CH
AD  - 2Center for Research in Biotechnology for Agriculture, University of Malaya, 
      50603 Kuala Lumpur, Malaysia. ISNI: 0000 0001 2308 5949. GRID: grid.10347.31
FAU - Kikuchi, Shinji
AU  - Kikuchi S
AD  - 1Laboratory of Genetics and Plant Breeding, Graduate School of Horticulture, 
      Chiba University, Matsudo, Chiba 271-8510 Japan. ISNI: 0000 0004 0370 1101. GRID: 
      grid.136304.3
FAU - Sassa, Hidenori
AU  - Sassa H
AD  - 1Laboratory of Genetics and Plant Breeding, Graduate School of Horticulture, 
      Chiba University, Matsudo, Chiba 271-8510 Japan. ISNI: 0000 0004 0370 1101. GRID: 
      grid.136304.3
FAU - Koba, Takato
AU  - Koba T
AD  - 1Laboratory of Genetics and Plant Breeding, Graduate School of Horticulture, 
      Chiba University, Matsudo, Chiba 271-8510 Japan. ISNI: 0000 0004 0370 1101. GRID: 
      grid.136304.3
LA  - eng
PT  - Journal Article
DEP - 20180510
PL  - England
TA  - Mol Cytogenet
JT  - Molecular cytogenetics
JID - 101317942
PMC - PMC5946395
OTO - NOTNLM
OT  - Abelia x grandiflora
OT  - Chloroform
OT  - Cucumis melo
OT  - FISH
OT  - Pachytene
OT  - Prometaphase
COIS- Not applicableThe authors declare that they have no competing interests.Springer 
      Nature remains neutral with regard to jurisdictional claims in published maps and 
      institutional affiliations.
EDAT- 2018/05/16 06:00
MHDA- 2018/05/16 06:01
PMCR- 2018/05/10
CRDT- 2018/05/16 06:00
PHST- 2018/02/20 00:00 [received]
PHST- 2018/04/23 00:00 [accepted]
PHST- 2018/05/16 06:00 [entrez]
PHST- 2018/05/16 06:00 [pubmed]
PHST- 2018/05/16 06:01 [medline]
PHST- 2018/05/10 00:00 [pmc-release]
AID - 380 [pii]
AID - 10.1186/s13039-018-0380-6 [doi]
PST - epublish
SO  - Mol Cytogenet. 2018 May 10;11:32. doi: 10.1186/s13039-018-0380-6. eCollection 
      2018.