Welcome to Chinese Journal of Tropical Crops,

Chinese Journal of Tropical Crops ›› 2020, Vol. 41 ›› Issue (8): 1522-1528.DOI: 10.3969/j.issn.1000-2561.2020.08.003

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Single-microspore Microdissection of Hevea brasiliensis

YANG Xingxing1,2,3,WANG Ying2,*(),ZHUANG Nansheng2,GAO Heqiong2,WANG Yuxiang2   

  1. 1. College of Forestry, Hainan University, Haikou, Hainan 570228, China
    2. College of Tropical Crops, Hainan University, Haikou, Hainan 570228, China
    3. Rubber Research Institute, Chinese Academy of Topical Agricultural Sciences, Danzhou, Hainan 571737, China
  • Received:2019-10-12 Revised:2020-01-28 Online:2020-08-25 Published:2020-09-17
  • Contact: WANG Ying


Hevea brasiliensis is an important tropical crop. In this study, the young male flowers of ‘Reyan 7-33-97’ in the meiotic tetrad period were used as the experimental materials to obtain single cells. Four enzymatic solutions, 5% cellulose, 5% pectinase, 5% β-glucanase, 5% cellulase and 4% pectinase mixed enzyme solution, were used in the experiment to explore the best scheme for the degradation of tetrad callose. As a result, under the condition of 1 h and 37℃, 5% cellulase and 4% pectinase mixed enzyme solution had the best enzymatic hydrolysis effect, and the single microspore with good quality could be obtained. Through the continuous exploration and experiment on the fine length of the needle tip, the diameter of the sharpening of the needle and the micro-operation separation system, a set of microdissection technology system suitable for the single microspore of ‘Reyan 7-33-97’ was successfully established. Single-microspores were the direct products of meiosis. The separation and analysis of single cells could accurately resolve gene recombination and exchange during the meiosis of rubber trees, and determine the genomic variation. The study would provide theoretical and technical support for single-cell whole genome sequencing of H. brasiliensis.

Key words: Hevea brasiliensis, microdissection, single cell, tetrad

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