不同培养条件对薄荷试管苗玻璃化现象的影响

以江苏东台产薄荷(M entha haplocalyxB riq.)的茎尖为外植体,以MS为基本培养基,研究了培养基添加物和培养条件对薄荷试管苗玻璃化现象的影响。结果显示,导致薄荷玻璃化苗产生的主要因素是培养基中的6-BA、蔗糖和琼脂浓度以及培养温度和光照时间;当6-BA浓度为2 mg.L-1、蔗糖浓度为4%、琼脂浓度为0.70%、培养温度25℃、光照12 h.d-1(2 000 lx)时,薄荷试管苗的繁殖系数较高,玻璃化苗率较低。     

甜瓜离体再生继代培养中玻璃化现象的研究

为提高甜瓜离体培养的再生率和转基因效率,以优质甜瓜品种‘伽师瓜’(‘卡拉库赛’)离体再生不定芽为外植体,通过连续多代继代培养,对引起玻璃化苗现象的几个主要因素进行了研究。结果表明,在甜瓜离体再生继代培养中,外植体继代次数是影响玻璃化发生的主要因素,同时培养基中的6-BA浓度偏高、琼脂浓度偏低以及蔗糖浓度偏低或偏高等可导致玻璃化苗的增加。培养基中较低的6-BA浓度(0~0.2 mg/L),琼脂浓度为6 g/L,蔗糖浓度为25 g/L以及添加活性炭等措施可有效地降低甜瓜玻璃化苗的发生。     

影响香石竹试管苗玻璃化的因素（简报）

香石竹组织培养中,当MS培养基中的6-BA浓度为0．5μg·ml－1,IAA浓度为0．2μg·ml－1,琼脂浓度为0．6％,光照时间为11h·d－1,温度在25．5℃左右并且通风状况良好时,香石竹试管苗玻璃化程度最低。     

油菜试管苗玻璃化发生机理的研究初探

研究了影响甘蓝型油菜离体下胚轴再生过程中试管苗玻璃化产生的几个因子,试验发现:调整激素6-BA与NAA的浓度、增加蔗糖的浓度提高渗透势对玻璃化无影响;降低培养基中NH4 浓度不仅没有有效地防止玻璃苗的发生反而降低了芽的诱导率;采用牛皮纸代替聚乙烯塑料膜作为封口材料的措施受到了地区的限制,没有得到有效的统计数据;选择1%的琼脂浓度可使玻璃化率显著降低55%,但诱芽率在一定程度上降低了6.9%。     

甜瓜试管苗继代培养玻璃化现象的研究

试管苗玻璃化现象是甜瓜组织基因转化过程中试管苗继代培养过程中的一大障碍,采用在培养基中提高糖和琼脂的浓度,添加适当的钙离子,细胞分裂素6-BA含量不超过0.2mg/L,生长素IAA用量控制在0.5mg/L,适当的光,光照强度和温度对克服甜瓜玻璃苗玻璃化现象有明显的效果.     

不同培养条件对光叶楮试管苗玻璃化的影响

以MS为基本培养基,研究生长调节剂、蔗糖、琼脂、光照强度和温度等因素对光叶楮试管苗玻璃化的影响。结果表明,最佳的生长调节剂配比是1.0 mg/L 6-BA、0.3 mg/L NAA,蔗糖浓度3.5%,琼脂浓度0.35%,光照强度2 000 lx,培养温度25℃。该优化培养条件能有效防止玻璃化苗的发生,获得较高的增殖系数和较健壮的苗木,且能降低生产成本。     

降低黄芩再生苗玻璃化率及其生根的研究

为了降低黄芩组培苗的玻璃化率并提高其生根率,本研究以黄芩无菌苗茎段诱导的不定芽为实验材料,分别研究了6-苄基嘌呤(6-BA)、蔗糖、琼脂及多效唑(PP333)等组培条件对黄芩不定芽玻璃化的影响以及吲哚丁酸(IBA)对再生苗生根的影响。研究结果表明:低浓度的6-BA有利于降低不定芽的玻璃化率,当培养基中添加0.2 mg·L~(-1)6-BA时,不定芽的玻璃化率最低且增殖系数比对照增加1倍。随着培养基中蔗糖浓度的升高,不定芽的玻璃化率显著降低,其增殖系数也有所降低。培养基中蔗糖浓度为25 g·L~(-1)时,黄芩不定芽长势最好且玻璃化率为零,增殖系数也最高。培养基中添加7.5 g·L~(-1)的琼脂,不定芽的玻璃化率较低,增殖系数也较高。培养基中添加多效唑(PP333)有利于缓解黄芩再生不定芽玻璃化状态,随着PP333的浓度增加,玻璃化率也逐渐降低。0.2 mg·L~(-1)PP333对黄芩不定芽的壮苗起到了很好的作用,再生苗明显变得粗壮。0.1 mg·L~(-1)的IBA最有利于黄芩再生苗的生根,生根率为100%,平均生根数为7,移栽成活率达95%以上。     

“固液双层”培养法诱导马铃薯‘米拉’试管薯的研究

该研究以马铃薯‘米拉’品种的脱毒试管苗为材料,采用"固液双层"的培养方式,通过正交试验对其试管苗壮苗生长阶段和试管薯诱导阶段的培养基进行优化,并通过单因素试验研究蔗糖浓度、光照条件和CCC浓度对试管薯结薯的影响。结果表明:在"固液双层"培养中,‘米拉’壮苗培养阶段优化的培养基为改良MS培养基(硝酸铵为2 000 mg·L~(-1)、硝酸钾为2 000 mg·L~(-1))+500 mg·L~(-1)CCC+0.1%活性炭+0.1mg·L~(-1)DA-6+1 mg·L~(-1)6-BA+0.1 mg·L~(-1)NAA+3%蔗糖+6 g·L~(-1)琼脂,pH 5.8。试管薯诱导及生长阶段优化的培养基为MS_1培养基(微量元素和铁盐的用量为MS培养基的2倍)+1.5%活性炭+4 mg·L~(-1)6-BA+8%蔗糖。在试管薯诱导阶段,弱光4 h·d~(-1)培养诱导的试管薯,结薯指数、大薯率、薯块重量均优于暗培养。"固液双层"培养是一种低成本的方法,在组织培养室内就可以大量繁殖‘米拉’试管薯,并且能增加原种的数量,这种方法也能用于马铃薯其他栽培品种试管薯的诱导。     

白萼吊钟海棠的组织培养与快速繁殖

以带节茎段为外植体进行了白萼吊钟海棠(Fuchsia alba-coccineaHort.)的组织培养和快速繁殖研究,对外植体的灭菌方法以及在试管苗增殖和生根培养过程中不同浓度的激素配比进行了筛选,同时研究了抑制试管苗褐化和玻璃化的方法。结果表明,最适宜白萼吊钟海棠外植体灭菌的方法是用0.1%HgC l2处理4~6 m in;MS培养基中不加NH4NO3可完全消除试管苗玻璃化现象;MS培养基中加入1.0 g.L-1PVP,可基本抑制试管苗褐化;试管苗增殖的最佳培养基为含0.8 mg.L-16-BA、0.10 mg.L-1NAA和1.0 g.L-1PVP的MS培养基;试管苗生根的最适培养基为含0.1~0.2 mg.L-1NAA和1.0 g.L-1PVP的1/2 MS培养基。     

培养条件对澳蜡花‘舞后’试管苗玻璃化的影响

以澳蜡花‘舞后’的试管苗为试验材料,研究了培养条件对其试管苗玻璃化的影响。结果表明,当以1/2MS为基本培养基,附加0.2mg·L-16-BA、0.01mg·L-1IBA、4%蔗糖、0.8%琼脂并在以塑料封口膜为封口材料的三角瓶内培养时,试管苗的增殖系数较高,玻璃化率较低。     

大花蕙兰组织培养技术研究

以大花蕙兰Cymbidium hybridum侧芽为外植体,以N6、B5、CC、MS为基本培养基,设计无机大量元素、无机微量元素、有机物三因素四水平正交试验,探究大花蕙兰丛生芽增殖的最佳基本培养基、生长调节剂浓度配比以及诱导生根的最佳生长调节剂浓度。结果显示,大花蕙兰丛生芽增殖的最佳培养基组合为B5无机大量元素+ CC无机微量元素+MS有机物+蔗糖30 g·L-1 +琼脂7 g·L-1 +活性炭0.5 g·L-1(pH 5.8～6.0),诱导丛生芽增殖的适宜生长调节剂浓度配比为6-BA 4.0 mg·L-1 + NAA 0.2 mg·L-1,诱导生根的最佳生长调节剂浓度为NAA 1.0 mg·L-1。     

‘香槟’月季的组织培养和试管开花诱导

本文对‘香槟’月季（80sachinensis‘Xiangbin’）的组织培养技术和诱导试管开花进行了研究。结果表明：以茎段为外植体能诱导获得无菌苗,适宜的启动培养基为MS＋6-BA1．0mg-L-1＋IBA0．1mg·L-1,幼芽继代增殖的最佳培养基是MS＋6．BA1．0mg·L-1。＋IBA0．1～0．2mg·L-1,诱导生根的适宜培养基为1／2MS＋NAA0．3mg·L-1,生根率达80．0％。诱导试管开花的适宜培养基为MS＋6．BA0．5mg·L-1＋NAA0．1mg·L-1最适宜的诱导试管开花的蔗糖含量是30g·L-1;在三角瓶中培养,试管花可以正常开放,在培养瓶中培养花芽不能正常开放;MS培养基中增加2倍磷的含量,可以提高花芽诱导率,为25．O％;诱导试管开花的最适培养条件为温度21℃,光照强度80～100μmol·m-2．s-1,光照时间16h—d-1。     

Effect of agar concentration on growth and anatomy of adventitious shoots of Picea abies (L.) Karst

The development of adventitious shoots of Picea abies was affected by the agar concentration in the culture medium. Increasing the agar concentration from 0.5 to 2.0% decreased vitrification, but at the same time reduced shoot growth and rooting potential. Slightly vitrified plantlets could become acclimatized to greenhouse conditions. The mesophyll of needles developed in vitro was interspersed with large air spaces; the lower the agar concentration, the larger the air spaces. After transfer to the greenhouse, the new needles from the acclimatized plantlets had an anatomy approaching that of plants growing in field.     

Cryopreservation of in vitro-grown shoot tips of apple and pear by vitrification

In vitro-grown shoot tips of apples (Malus domestica Borkh. cv. Fuji) were successfully cryopreserved by vitrification. Three-week-old in vitro apple plantlets were cold-hardened at 5°C for 3 weeks. Excised shoot tips from hardened plantlets were precultured on a solidified Murashige & Skoog agar medium (MS) supplemented with 0.7 M sucrose for 1 day at 5°C. Following preculture shoot tips were transferred to a 2 ml plastic cryotube and a highly concentrated cryoprotective solution (designated PVS2) was then added at 25°C. The PVS2 contains (W/V) 30% glycerol, 15% ethylene glycol and 15% dimethylsulfoxide in medium containing 0.4 M sucrose. After dehydration at 25°C for 80 min, the shoot tips were directly plunged into liquid nitrogen. After rapid warming, the shoot tips were expelled into 2 ml of MS medium containing 1.2 M sucrose and then plated on agar MS medium. Direct shoot elongation was observed in approximately 3 weeks. The average rate of shoot formation was about 80%. This vitrification method was successfully applied to five apple species or cultivars and eight pear cultivars. This method appears to be a promising technique for cryopreserving shoot tips from in vitro-grown plantlets of fruit trees.Abbreviations DMSO dimethylsulfoxide - EG ethylene glycol - PVS2 vitrification solution - LN liquid nitrogen - BA 6-benzylaminopurine - NAA -naphthaleneacetic acid - SE standard error - ABA abscisic acid     

Effects of agar brand and concentration on the tissue culture medium

In tissue-cultured Cynara scolymus L. (globe artichoke) vitrification (hyperhydric transformation) can only be overcome by increasing the concentration of agar. However, with increasing concentrations of Difco Bacto agar (8–15 g/I) the availability of labelled kinetin is decreased. There is some evidence for postulating that cytokinins under inductive conditions of low agar concentration or high matrix potential are evocators of vitrification.
Both the brand and concentration of agar also affect the chemical and physical characteristics of a culture medium.
Impurities introduced with the agar are responsible for significant differences in the concentration of an element in comparable media with different levels of agar, and are easily detected by conductance.
Penetrometer measurements also show large ranges in solidity of media with increasing concentrations, not only within the type of agar, but also for the same concentration within different brands.     

风信子花被外植体年龄对花器官分化的影响

离体培养风信子(Hyacinthus orientalis L．)不同年龄的花被外植体诱导花器官直接再生的实验表明;1．在MS附加6-BA 2 mg／L,2,4-D 0.1 mg／L的培养基上,年龄V的外植体大量衰老,基本丧失器官再生能力,处于年龄段Ⅱ～Ⅳ的外植体可发生玻璃化反应。2．玻璃化反应的外植体转移至MS附加6-BA 0.2 mg／L、NAA 0.005 mg／L的培养基上继续培养30 d后可再生正常的花被片,表明降低培养基中外源激素浓度能够阻止玻璃化反应继续发生。3．在MS附加6-BA 2 mg／L．2,4-D 0.1 mg／L的培养基上,外植体形态学下部可再生雌蕊状早期结构。平均每块外植体分化雌蕊状早期结构数以年龄Ⅲ的外植体最多。     

中国石竹离体快繁与试管苗玻璃化研究

研究了培养基组成成分对中国石竹品种Diana F1 White组培程序中种子萌发、诱芽增殖、试管苗玻璃化及生根等重要环节的影响。结果表明:(1)种子适宜的萌发培养基为1/2MS,在此培养基中种子萌发率为83.33%,播种后30d时无菌苗高度达6.99cm,叶片数达26.7。(2)在芽诱导阶段玻璃化苗发生率最高可达53.85%。将光照强度提高至2000lx后,采用培养基MS+6-BA3.0mg/L+NAA0.3mg/L+琼脂8.0g/L+蔗糖8.0g/L进行诱芽培养,玻璃化苗发生率降至3.33%,外植体出芽率达到90%,单个外植体出芽数达到7.2个。(3)适宜的生根培养基为1/2MS+1.0mg/LIBA+琼脂8g/L+蔗糖40g/L,培养30d时生根率为100%,平均根条数达到24.7条,平均根长度达到4.7cm。试管苗在消毒后的腐殖土中移栽,成活率达95%。该研究结果为DianaF1试管苗的快速繁殖提供科学依据。