吉林师范大学学报（自然科学版）
吉林師範大學學報（自然科學版）
길림사범대학학보（자연과학판）
JILIN NORMAL UNIVERSITY JOURNAL（NATURAL SCIENCE EDITION）
2009年
3期
61-65
,共5页
高山红景天%茎尖%包埋-玻璃化法%超低温保存
高山紅景天%莖尖%包埋-玻璃化法%超低溫保存
고산홍경천%경첨%포매-파리화법%초저온보존
Ph.sachalinensis%shoot tips%encapsulation-vitrification%cryopreservation
采用包埋.玻璃化法对高山红景天茎尖进行了超低温保存研究,研究了蔗糖预处理、包埋过程中渗透和不同温度下玻璃化液PVS2处理的时间对超低温保存后茎尖存活率的影响.结果表明,茎尖在0.3,0.5,0.7,1.0mol·L-1的蔗糖溶液中各预培养1 d后,转入1.0 mol·L-1的蔗糖溶液中再培养4 d,使用含2 mol·L-1甘油和1mol·L-1蔗糖的包埋液渗透处理60 min,包埋珠在0℃处理180至210分钟后投入液氮,投入48 h后用40℃水浴快速化冻3 min,用含有1 mol·L-1蔗糖的改良MS培养液洗涤20 min,转入MS+6-BA 1mg·L-1+NAA0.1 mg·L-1固体培养基中在22℃条件下进行暗培养,2 d后转入光照培养,最高成活率接近100%,再生植株生长和分化正常.
採用包埋.玻璃化法對高山紅景天莖尖進行瞭超低溫保存研究,研究瞭蔗糖預處理、包埋過程中滲透和不同溫度下玻璃化液PVS2處理的時間對超低溫保存後莖尖存活率的影響.結果錶明,莖尖在0.3,0.5,0.7,1.0mol·L-1的蔗糖溶液中各預培養1 d後,轉入1.0 mol·L-1的蔗糖溶液中再培養4 d,使用含2 mol·L-1甘油和1mol·L-1蔗糖的包埋液滲透處理60 min,包埋珠在0℃處理180至210分鐘後投入液氮,投入48 h後用40℃水浴快速化凍3 min,用含有1 mol·L-1蔗糖的改良MS培養液洗滌20 min,轉入MS+6-BA 1mg·L-1+NAA0.1 mg·L-1固體培養基中在22℃條件下進行暗培養,2 d後轉入光照培養,最高成活率接近100%,再生植株生長和分化正常.
채용포매.파리화법대고산홍경천경첨진행료초저온보존연구,연구료자당예처리、포매과정중삼투화불동온도하파리화액PVS2처리적시간대초저온보존후경첨존활솔적영향.결과표명,경첨재0.3,0.5,0.7,1.0mol·L-1적자당용액중각예배양1 d후,전입1.0 mol·L-1적자당용액중재배양4 d,사용함2 mol·L-1감유화1mol·L-1자당적포매액삼투처리60 min,포매주재0℃처리180지210분종후투입액담,투입48 h후용40℃수욕쾌속화동3 min,용함유1 mol·L-1자당적개량MS배양액세조20 min,전입MS+6-BA 1mg·L-1+NAA0.1 mg·L-1고체배양기중재22℃조건하진행암배양,2 d후전입광조배양,최고성활솔접근100%,재생식주생장화분화정상.
Shoot tips excised from two resources of Rh. sachalinensis had been cryopreserved by encapsulation- vitrification. In the experiment, effects of sucrose preculture duration, osmoprotection time during encapsulation and exposure time to PVS2 on survival rate of cryopreserved shoot tips of Rh. sachalinensis were discussed. Mature seeds of Rh. sachalinensis were cultured on hormone-free MS medium,and plantlets formed from germinated seeds in 3 weeks. Excised shoot tips were precultured with increasing sucrose concentrations of 0.3,0.5,0.75 and 1 M for 4 days,then transferred to 1 M sucrose for 4 days, and precultured shoot tips were encapsulated and simultaneously osmoprotected with a loading solution of 2 M glycerol and 1 M sucrose for 60 rain. Osmoprotected and encapsulated shoot tips were dehydrated with a highly concentrated vitrification solution for 180 to 210 min at 4 ℃ prior to direct immersion in liquid nitrogen for 48 h. After rapid warming in water bath for 3 min at 40 ℃, the shoot tips were rinsed with the modified MS medium containing 1 M sucrose for 20 min,and then cultured on the MS medium supplemented with 6-BA 4.44 μM and NAA 0.54 μM in dark for 1 week prior to exposure to the light at 22℃. The best survival rate of encapsulation vitrified shoot tips amounted to nearly 100% and no planflet abnormality was observed.
