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园艺学报 ›› 2020, Vol. 47 ›› Issue (2): 301-309.doi: 10.16420/j.issn.0513-353x.2020-0195

• 研究论文 • 上一篇    下一篇

朱顶红无菌苗叶片高效再生体系

王春夏,张梦迪,王锦霞,王志平,孙红梅   

  1. 沈阳农业大学园艺学院,设施园艺省部共建教育部重点实验室,北方园艺设施设计与应用技术国家地方联合工程研究中心,沈阳 110866
  • 出版日期:2020-02-25 发布日期:2020-02-25
  • 基金资助:
    辽宁省科技攻关重大项目(201015003)

Establishment of an Efficient Regeneration System in Hippeastrum vittatum with Plantlet Leaves

WANG Chunxia,ZHANG Mengdi,WANG Jinxia,WANG Zhiping,and SUN Hongmei   

  1. College of Horticulture,Shenyang Agricultural University;Key Laboratory of Protected Horticulture of Education Ministry and Liaoning Province;National & Local Joint Engineering Research Center of Northern Horticultural Facilities Design & Application Technology,Shenyang 110866,China
  • Online:2020-02-25 Published:2020-02-25

摘要: 以朱顶红(Hippeastrum vittatum)叶片为外植体进行离体培养,具有取材方便、试材充足、成本低等优势,但叶片诱导再生率极低,是朱顶红离体培养的一大难题。本试验中分别以‘花孔雀’和‘黑天鹅’朱顶红无菌苗叶片为外植体,探究了不同植物生长调节剂和不同取材部位对不定芽诱导和继代增殖的影响。结果表明:最佳外植体为 MS 培养基中培养 10 d 形成的幼嫩叶片基部(0.5 cm),在光照 16 h · d-1(光照强度 36 μmol · m-2 · s-1)下,不定芽诱导的最适培养基为 MS + 2 mg · L-1 6-BA + 1 mg · L-1 NAA + 2 mg · L-1 TDZ,两个品种的不定芽均以间接途径发生,其中‘花孔雀’在培养 40 d 后形成愈伤组织,55 d形成不定芽,诱导率可达 69.44%;‘黑天鹅’在培养 45 d 后形成愈伤组织,65 d 形成不定芽,诱导率达到 66.67%;最适体细胞胚诱导培养基为 MS + 2 mg · L-1 6-BA + 1 mg · L-1 PIC,‘花孔雀’和‘黑天鹅’的诱导率分别达到 66.67%和 63.89%;最佳不定芽增殖培养基为 MS + 2 mg · L-1 6-BA + 1 mg · L-1 NAA + 1 mg · L-1 TDZ,‘花孔雀’和‘黑天鹅’的增殖系数分别达到 4.67 和 3.46;在不添加植物生长调节剂的 MS培养基中进行生根培养,30 d 后两个品种的生根率均达到 100%;将生根培养 30 d 的小植株转移至室温条件下放置 3 d,摘去封口膜再驯化 3 d 后,移栽至经高温消毒的草炭︰蛭石(体积比)为 1︰1 的基质中,成活率达到 100%

关键词: 朱顶红, 离体培养, 高效再生, 无菌苗叶片

Abstract: Using the leaves as explants in vitro culture,it has the advantages of convenient materials,abundant test materials,and low cost. However,the induced regeneration rate of leaves is extremely low,which is a major problem in vitro culture of Hippeastrum vittatum. Effects about different plant growth regulators and different parts of materials on adventitious bud induction and subculture were studied with the leaves of‘Blossom Peacock’and‘Royal Velvet’plantlet. The results showed that the best explants were the base of young leaves(0.5 cm)formed by culturing in MS medium for 10 days. The photoperoid is 16 h · d-1(light intensity 36 μmol · m-2· s-1). The best medium for adventitious buds induction of Hippeastrum vittatum was MS + 2 mg · L-1 6-BA + 1 mg · L-1 NAA + 2 mg · L-1 TDZ. The adventitious buds of both varieties occurred in an indirect way,and the‘Blossom Peacock’formed callus after 40 days of culture,adventitious buds formed on 55 days,the induction rate reached 69.44%;‘Royal Velvet’formed callus after 45 days of culture,and adventitious buds formed on 65 days,the induction rate reached 66.67%. The optimal adventitious bud proliferation medium was MS + 2 mg · L-1 6-BA + 1 mg · L-1 NAA + 1 mg · L-1 TDZ,and the proliferation coefficients of‘Blossom Peacock’and‘Royal Velvet’reached 4.67 and 3.46,respectively. The optimal somatic embryo induction medium was MS + 2 mg · L-1 6-BA + 1 mg · L-1 PIC,and the induction rate of‘Blossom Peacock’and‘Royal Velvet’reached 66.67% and 63.89%,respectively. Rooting culture was carried out in MS medium without added hormones,and the rooting rate of both varieties reached 100% after 30 days. The Hippeastrum vittatum plantlets after 30 days of rooting culture were moved to room temperature for 3 days. Then remove the sealing film domestication for 3 days. They were transplanted to the high temperature sterilized charcoal︰vermiculite(volume ratio) 1︰1 matrix,and then the survival rate reached 100%.

Key words: Hippeastrum vittatum, in vitro culture, efficient regeneration, plantlet leaves

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