Autopolyploid induction via somatic embryogenesis in Lilium distichum Nakai and Lilium cernuum Komar
Autopolyploid induction via somatic embryogenesis in Lilium distichum Nakai and Lilium cernuum Komar
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通过体细胞胚胎发生诱导百合 Nakai 和百合 Komar 的同多倍体
DOI：
10.1007/s11240-019-01671-x
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发表时间：
2019-11-01
影响因子：
3
通讯作者：
Sun, Hongmei
中科院分区：
文献类型：
--
作者：
Fu, Linlan;Zhu, Yingying;Sun, Hongmei
通过人工多倍体诱导可以创造出高品质的观赏新品种。本研究首次建立了百合和东方百合体细胞胚胎发生的多倍体诱导体系。是发展起来的。体细胞胚胎培养在MS+0.41mmolL-1NAA+1.07mmolL-−-1NAA5 mm~2的MS上。40d后，将体细胞胚胎转移到含有2.21μ摩尔的L−1 BA的MS上进行体细胞胚胎发生。石蜡切片观察表明，二节花的胚性细胞最初起源于外细胞，体细胞胚胎发生是通过间接途径进行的。在羊草中，胚胎细胞最初起源于内细胞，体细胞胚胎发生是通过直接途径进行的。采用不同秋水仙素浓度(0.01%、0.05%和0.1%；v/v)和培养时间(24、48、72h)的浸泡和混合培养方法，成功地从体细胞胚胎和鳞片中获得了多倍体。0.05%秋水仙素处理二节草48 h和小叶24 h，多倍体诱导率分别达到57.14%和46.15%。在获得的多倍体试管苗中，通过根尖组织压片染色体计数鉴定出四倍体(28.57%)和非整倍体(23.08%)。与二倍体植株相比，四倍体植株叶片较宽，保卫细胞较长，气孔较大，气孔导度较高。四倍体植株的叶绿素含量比二倍体植株高1.76 × ，叶片数显著增加，保卫细胞较长，气孔较大，气孔导度较低。本研究首次建立了利用体细胞胚胎和鳞片浸泡和混合培养的方法，建立了二枝莲和羊草的体细胞胚胎发生体系。比较秋水仙碱浓度和时间的组合，以选择最佳治疗组合。采用形态观察、气孔观察和根尖组织压片相结合的方法鉴定加倍植株的倍性。本研究结果为提高两种野生百合的观赏价值，创造新的百合种质资源，提高资源的繁殖系数提供了依据。
New ornamental varieties of high quality can be created via artificial polyploid induction. In the present study, the first system of polyploid induction with somatic embryogenesis of Lilium distichum Nakai and Lilium cernuum Komar. was developed. Somatic embryos were cultured on MS with 0.41 μmol L−1 picloram and 1.07 μmol L−1 NAA by scales (5 mm2). After 40 days, somatic embryos were transferred to MS with 2.21 μmol L−1 BA for somatic embryogenesis. As determined from observations of paraffin sections, embryonic cells of L. distichum originated from outer cells at first, and somatic embryogenesis occurred through an indirect pathway. In L. cernuum, embryonic cells originated from inner cells at first, and somatic embryogenesis occurred through a direct pathway. Polyploids were successfully formed from somatic embryos and scales by the soaking and mixed culture methods with different colchicine concentrations (0.01%, 0.05%, and 0.1%; v/v) and durations (24, 48, and 72 h). The polyploid induction rate reached 57.14% and 46.15% with 0.05% colchicine treatment in L. distichum (48 h) and L. cernuum (24 h), respectively. Tetraploids (28.57% and 23.08%) and aneuploids without chimeras among the obtained polyploid plantlets were identified by chromosome counts of root-tip tissue squashes in L. distichum and L. cernuum. Tetraploid plantlets of L. distichum exhibited broader leaves, longer guard cells, larger stomata and higher stomatal conductance than diploid plantlets. Tetraploid plantlets of L. cernuum showed 1.76 × higher chlorophyll content, significantly more leaves, longer guard cells, larger stomata and lower stomatal conductance than diploid plantlets. For the first time, we established a somatic embryogenesis system for L. distichum and L. cernuum using somatic embryos and scales to induce polyploids by soaking and mixed culture. Combinations of colchicine concentrations and periods of time were compared to select the best treatment combination. Comprehensive morphological observations, stomatal observations and root-tip tissue squashes were used to identify the ploidy of doubling plants. Our results provide a foundation for improving the ornamental value of two wild lily species, creating new Lilium germplasm and improving the reproduction coefficient of these resources.