摘要
研究了魔芋茎尖玻璃化冻存的主要技术环节，建立了较适宜的魔芋种质资源超低温保存体 系：在实体显微镜下，从继代培养 ２周的不定芽上切取 １～１． ３３ ｍｍ大小的茎尖，经０． ７ ｍｏｌ／Ｌ蔗糖的 ＭＳ－ＮＨ４＋培养液室内预培养 ２ｄ，玻璃化溶液ＰＶＳ２室温下处理 １０～２０ ｍｉｎ，直接投入液氮中保存。保 存后，茎尖经 ４０～７７℃水浴快速化冻， １． ２ ｍｏｌ／Ｌ蔗糖的培养液洗 ２次，各 １０ ｍｉｎ，暗中培养 ２周后转 入光下。冻存的茎尖直接生长发育或经愈伤组织再分化形成完整植株，存活率为５０％～７０％。从基因 组ＤＮＡ检测结果表明，魔芋茎尖经超低温保存后，其遗传特性未发生改变。
The factors affecting shoot tips cryopreservation of Amorphophallus, such as preculture, treatment with PVS2, and thawing temperature were investigated. A suitable procedure was established as follows: two weeks after subculture, shoot tips (1 - 1. 33 mm long) were taken from buds which were growing and developing quickly and were precultured on MS-NH4+ medium supplemented with 0. 7 mol/L sucrose for 2 days at 25℃. Then they were treated with a vitrification solution (PVS2) at 25℃ for 10-20 min prior to a direct plunge into liquid nitrogen. After rapid warming in 40-77℃ water bath, shoot tips were expelled into MSNH4+ medium containing 1. 2 mol/L sucrose for 20 min and plated on solidified MS-NH4+ medium. The cryopreserved materials were cultured in darkness for 2 weeks so that they could grow quickly. Plantlets were formed from calli or shoots within 4 weeks. The average survival rates were from 50% to 70%. The genetic integrity of cryopreserved shoot tips was identified through products of PCR with arbitrary primers in genomic DNA from treated and untreated materials and no changes were found.
出处
《作物学报》
CAS
CSCD
北大核心
2001年第1期97-102,共6页
Acta Agronomica Sinica
基金
国家自然科学基金项目!(30670513)
博士学科点基金项目!(950605)资助
