红花檵木花粉单倍体愈伤组织的诱导与培养
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国家自然科学基金:31071826;湖南省自然基金:04JJ3024;湖南省教育厅省高等学校产业化培育项目:07CY001


Induction and cultivation of haploid callus of Loropetalum chinense var. rubrum
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    摘要:

    为通过红花檵木单倍体愈伤组织获得红花檵木全基因组序列,在红花檵木‘细叶玫红’减数分裂和小孢子发育进程研究的基础上,初步明确了与小孢子不同发育时期相对应的花蕾外部特征,筛选出‘细叶玫红’单核靠边期花粉进行花药愈伤组织培养。诱导培养基以SNGM为基本培养基,添加2.5 mg/L NAA +0.5 mg/L 6–BA+30 g/L蔗糖;花粉愈伤组织培养的最优培养基为B5培养基,添加物同诱导培养基;花药愈伤组织悬浮培养体系为B5液体培养基,添加物同诱导培养基,转速为110 r/min,培养温度为25 ℃。经流式细胞仪检测,红花檵木花粉愈伤组织最大吸收峰值约为叶片愈伤组织的50%;用染色体压片计数法观察到叶片愈伤组织细胞染色体数为24条,花粉愈伤组织为12条,单倍体细胞占63.1%,表明红花檵木‘细叶玫红’花粉经诱导培养出的愈伤组织为单倍体愈伤组织。

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    For obtaining the complete genome sequence of Loropetalum chinense var. rubrum, haploid callus was induced and cultivated based on previous data of meiosis and microspore development of Loropetalum chinense var. rubrum ‘xiyemeihong’. The outer characteristics of the flower bud corresponding to specific development stages were primarily determined and monocaryotic phase of ‘xiyemeihong’ was selected for induction of haploid callus. SNGM medium supplemented with 2.5 mg/L NAA, 0.5 mg/L 6–Benzyladenine and sucrose concentration 30 g/L turned out to be the optimum callus-induction medium. And B5 medium with supplements the same to the induction medium was the optimum medium for semi-solid-state cultivation of pollen callus and for liquid cultivation of anther callus under the rotation speed of 110 r/min and the temperature of 25 ℃. Flow cytometry showed that the absorption peak of the pollen callus was about half of that of the leave callus (the control). Chromosome counting indicated haploids of pollen callus possessed 12 chromosomes while diploids 24 and haploids were detected in 63.1% of pollen callus cells.

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李炎林,于晓英,熊兴耀,陈己任,朱杰辉,李达.红花檵木花粉单倍体愈伤组织的诱导与培养[J].湖南农业大学学报:自然科学版,2011,37(6):632-636.

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  • 收稿日期:2011-04-26
  • 最后修改日期:2011-05-23
  • 录用日期:2011-08-29
  • 在线发布日期: 2011-12-26
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