用RAPD法研究火炬松愈伤组织发生的遗传特性(英文)

体外培养对于植物的快速繁殖是非常有效的。和其它一些松果类硬木植物一样，火炬松的体外培养成功率却一直很低。本工作研究了不同的基本培养基和低温条件对于火炬松Ｊ－５６， Ｓ－１００３， ａｎｄ Ｅ－４４０等三个品系的成熟合子胚形成愈伤组织、分化出芽、成苗的影响。在不同的基本培养基条件下芽分化的程度差异很大。合子胚经过９－１２周培养，开始分化，形成具有器官发生的愈伤组织（Ｆｉｇ．２ａ）。分化后３周，开始诱导出芽（Ｆｉｇ．２ｂ），芽的生长快慢不同（Ｆｉｇ．２ｃ，ｄ）。同一个愈伤组织上会生出几个芽来（Ｆｉｇ．２ｅ）。在添加有ＩＢＡ和ＢＡ的ＴＥ培养基上芽生长最快（Ｆｉｇ．１）。低温条件持续 １５天，能增加芽的数量和分化的程度（Ｔａｂｌｅ１）。上述培养基中增加ＧＡ３时表明，ＧＡ３对于根的诱导有决定性的作用。将９８株再生苗转移到特殊的混合土壤上；成活了７５株苗（Ｆｉｇ．２ｆ）。以这三种火炬松的再生苗尖为材料制备ＤＮＡ。用２０个引物进行ＲＡＰＤ分析，结果表明：这三种火炬松苗的扩增产物是相同的（Ｆｉｇ．２ｇ，ｈ＆ｉ）。这说明：用愈伤组织克隆植株的过程中没有引起植物遗传变异。

Plantlets using Random Amplified Polymorphic DNA (RAPD) Diagnosis

Adventitious buds were induced from organogenic callus derived from mature zygotic embryos of three families (J-56, S-1003, and E440) of loblolly pine (Pinus taeda L.) within 9-12 weeks of culture. Influences of basic media and low temperature treatment on the differentiation of adventitious buds were analyzed. Elongation of adventitious buds was achieved on TE medium supplemented with 0.5 mg/L indole-3-butyric acid (IBA) and I mg/L 6-benzyladenine (BA). After the adventitious shoots rooted on TE medium supplemented with 0.5 mg/L IBA, 2 mg/L BA, and 0.5 mg/L gibberellic acid (GA3), 98 regenerated plantlets were transferred to a perlite:peatmoss:vermiculite (1:1:1) soil mixture and 75 plantlets survived in the field. Total DNA was extracted from the needles of the regenerated plantlets of the three families of loblolly pine. Analysis of random amplified polymorphic DNA (RAPD) using 20 arbitrary oligonucleotide 10-iners showed that the amplification products were monomorphic for all the plantlets deviled from the families, J-56, S-1003, and E-440, of loblolly pine. These results suggested that the somatic organogenesis could be used for clonal micropropagation of some families of loblolly pine.