Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (17): 3729-3735.doi: 10.3864/j.issn.0578-1752.2013.17.023

• RESEARCH NOTES • Previous Articles     Next Articles

Exploration and Application of Non-Denaturing FISH on Chromosomes Research in Sesame (Sesamum indicum L.)

 LIU  Yan-Yang, CUI  Cheng-Qi, MEI  Hong-Xian, WU  Ke, ZHENG  Yong-Zhan   

  1. Henan Academy of Agricultural Sciences/Henan Sesame Research Center, Zhengzhou 450002
  • Received:2013-04-10 Online:2013-09-01 Published:2013-07-05

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Abstract: 【Objective】The objective of this paper is to study the optimal experimental conditions for non-denaturing FISH (ND-FISH), and to explore its application for chromosome recognition in sesame. 【Method】 Using oligomeric nucleotide (AC)8 as probe, the effects of different elution strengths, digestions and hybriding times on the ND-FISH were analyzed to optimize the protocol. 【Result】The optimal experimental conditions for ND-FISH were as follows: cells on slides were treated with DNase-free RNase A at a concentration of 100 μg•mL-1 for 1 h, 6 µL of the probe (20 ng•µL-1), hybridization for 4 h, elution with the 4×SSC/0.2% Tween 20 optimal solution for 10 min. Using (AC)8 as a probe, mitotic metaphase chromosomes of Yuzhi 11 were analyzed by ND-FISH. Nine pairs of the chromosomes of Yuzhi 11(seven pairs of chromosomes had strong signals, and two pairs of chromosomes had weak signals) could be distinguished based on the hybridization signals combining with the length and arm ratio of chromosomes. 【Conclusion】ND-FISH technology with (AC)8 as a probe was used to successfully identify 18 of 26 chromosomes of cultivated sesame for the first time.

Key words: sesame , oligonucleotide , chromosome , karyotype , ND-FISH

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