红掌特伦萨四倍体的离体诱导及其鉴定

Induction and Characterization of Anthurium andraeanum Turenza Tetraploids in Vitro

为了研究红掌四倍体的离体诱导和鉴定技术体系,为其倍性育种和杂交育种奠定基础,以特伦萨愈伤组织为试验材料,采用0.5、1.0、2.0、4.0 g·L^(-1)的秋水仙素分别处理2、4、6、8 h,之后转入培养基MS+6-BA 2.0 mg·L^(-1)+NAA 0.5 mg·L^(-1)进行愈伤组织的生长和丛生芽分化,75 d后将丛生芽转入培养基1/2 MS+NAA 0.2 mg·L^(-1)诱导生根,待丛生芽生根后统计愈伤组织块存活率、再生植株数,并对再生植株进行染色体倍性鉴定和核型分析。结果表明,随着秋水仙素浓度的增加,红掌愈伤组织块存活率呈下降趋势,但愈伤组织块上诱导的再生植株数无显著变化;在0.5~2.0 g·L^(-1)的秋水仙素浓度范围内,红掌疑似四倍体的诱导率随着处理浓度与时间的增加而增加,2.0 g·L^(-1)秋水仙素处理6与8 h时,诱导率达8.50%与8.77%;特伦萨和疑似四倍体红掌的基因组分别为3.73和7.01 Gb;染色体压片倍性鉴定结果证明了疑似四倍体的倍性为四倍体。红掌二倍体的核型公式为2n=2x=30=8 st+16 sm+6 m,四倍体的核型公式为2n=4x=60=8 st+32 sm+20 m;与红掌二倍体植株相比,四倍体植株叶片变厚、气孔变大、保卫细胞密度变小,佛焰苞直径变大,单个花序花期变长,差异均达显著水平。综上,采用2.0 g·L^(-1)秋水仙素处理6 h可培育红掌四倍体植株,此四倍体可作为红掌倍性育种和杂交育种的重要种质资源。

This study aims to establish the technical system of in vitro induction and identification ofAnthurium andraeanum tetraploids, laying the foundation for ploidy breeding and hybrid breeding. Trenzacalluses were used as materials to be treated with 0. 5, 1. 0, 2. 0 and 4. 0 g·L^(−1) of colchicine for 2, 4, 6 and8 h, respectively. Afterwards, the calluses were transferred to medium MS + 6-BA 2. 0 mg·L^(−1)+NAA0. 5 mg·L^(−1) for growth and clumped shoot differentiation. After 75 d, the clumped shoots were transferred tomedium 1/2 MS+NAA 0. 2 mg·L^(−1) to induce rooting. Subsequently, the survival rate of the callus blocks andthe number of regenerated plants were counted, and chromosome ploidy identification and karyotype analysiswere performed on the regenerated plants. The results showed that with the increase of colchicineconcentration, the survival rate of Anthurium andraeanum callus block showed a decreasing trend, but therewas no significant change in the number of regenerated plants induced on the callus block. Within theconcentration range of 0. 5 to 2. 0 g·L^(−1), the induction rate of suspected tetraploid of the Anthurium andraeanumincreased with the increase of the treatment concentration and the time, and the induction rate reached 8. 50%and 8. 77% when treated with 2. 0 g·L^(−1) colchicine for 6 and 8 h. The genomes of Turenza and suspectedtetraploid Anthurium andraeanum were 3. 73 and 7. 01 Gb, respectively. The results of chromosome ploidyidentification confirmed that the suspected tetraploid was indeed tetraploid. The karyotype formula forAnthurium andraeanum diploid was 2n=2x=30=8 st+16 sm+6 m and that for tetraploid was 2n=4x=60=8 st+32 sm+20 m. Significant differences were observed in thicker leaf blades, larger stomata, lower density ofguard cells, larger spathe diameters and longer flowering period of individual inflorescences in tetraploidplants as compared to Anthurium andraeanum diploid plants. In summary, treatment with 2. 0 g·L^(−1) colchicinefor 6 h can cultivate tetraploid plants of Anthurium andraeanum, which can be serve as an importantgermplasm resource for ploidy breeding and cross breeding of Anthurium andraeanum.