不同浓度卡那霉素、潮霉素对楸树试管苗生长的影响

目的:将不同质量浓度的卡那霉素、潮霉素加入楸树培养基中,研究卡那霉素、潮霉素对楸树组培苗生长的影响,以确定抗生素对楸树茎段分化与生根的敏感质量浓度。方法:待楸树继代、生根培养基灭菌后温度降至30~50℃,将不同质量浓度的卡那霉素、潮霉素经抽滤式灭菌加入培养基中,在培养基中接入楸树组培无菌茎段培养,观测茎段继代(增殖芽数、芽长、叶数等)、生根(发根数、根长、芽长等)生长指标并统计分析。结果:楸树组培继代培养基添加卡那霉素质量浓度为100 mg/L时组培瓶苗生长缓慢,浓度为150 mg/L时叶片大部分发白并干枯,茎段基部无愈伤组织形成,瓶苗基本停止生长,楸树继代瓶苗对卡那霉素耐受性范围为100~150 mg/L;添加潮霉素质量浓度为5 mg/L时瓶苗生长较为缓慢,浓度为10 mg/L时叶片开始干枯,茎段基部愈伤组织较小,瓶苗基本停止生长,楸树继代瓶苗对潮霉素耐受性范围为10 mg/L左右。楸树组培生根培养基添加卡那霉素质量浓度为100 mg/L时大部分茎段干枯,少部分为绿但未分化芽与根,浓度为150 mg/L时大部分茎段干枯,极少上部为绿,基部干枯,但未分化芽与根,楸树组培瓶苗生根培养苗对卡那霉素耐受性范围为100~150 mg/L;添加潮霉素质量浓度为5 mg/L时少部分茎段干枯,浓度为10 mg/L时大部分茎段干枯,少部分为绿,茎段未出现芽的分化与根的萌发现象,楸树组培瓶苗生根培养苗对潮霉素耐受性范围为5~10 mg/L。结论:卡那霉素、潮霉素对楸树组培苗生长有明显的抑制作用且与抗生素浓度呈负相关,但低质量浓度(1 mg/L)的潮霉素对楸树继代分化芽数有促进作用;同一抗生素对楸树不同无性系间组培苗生长的影响无显著差异。     

大花蕙兰组培快繁技术研究

选用大花蕙兰杂交新品种1053为外植体,采用1/2MS为基本培养基进行大花蕙兰组培快繁技术研究,结果表明：类原球茎诱导的最佳外植体为大花蕙兰鳞茎,培养基最佳激素浓度配比为0.5 mg/L 6-BA＋0.2 mg/L NAA;类原球茎增殖与分化芽最佳激素浓度配比为1.0 mg/L 6-BA＋0.3 mg/L NAA;幼芽增殖的最佳激素浓度配比为2.0 mg/L 6-BA＋0.5 mg/L NAA,最佳有机添加物为150 ml/L椰汁,幼芽增殖正交试验得出影响因素主次顺序：6-BA〉椰汁添加物〉NAA,优组合为2 mg/L 6-BA、0.5 mg/L NAA、添加物椰汁150 ml/L;诱导生根的最佳激素浓度配比为0.3 mg/L IBA＋0.2 mg/L NAA,添加香蕉泥100 g/L对幼苗根生长有显著的促进作用。优化后的大花蕙兰组培快繁技术参数,可为大规模工厂化生产提供参考。     

文心兰原球茎液体增殖培养研究

以茎尖诱导形成的原球茎(protocorm-like bodies,PLBs)为外殖体,采用液体培养方式比较了不同浓度的激素配比、蔗糖浓度和添加不同量的新鲜椰汁对文心兰PLBs增殖的影响,并比较了相同培养基成分时液体培养PLBs增殖、分化成苗和固体培养PLBs增殖和分化成苗的差异。试验结果表明:不同浓度的外源激素及其配比对文心兰PLBs增殖生长影响较大,6-BA0.5 mg/L Ad 0.05 mg/L NAA0.05 mg/L的激素组合比较适合文心兰PLBs增殖;蔗糖浓度对文心兰PLBs增殖的影响也较大,适合文心兰PLB在液体培养条件下增殖的蔗糖浓度为7.5 g/L;添加5%新鲜椰汁不仅对文心兰PLBs增殖有促进作用,而且能改善PLBs的质量;适合文心兰PLBs增殖的培养基为MS 6-BA0.5 mg/L Ad 0.05 mg/L NAA0.05 mg/L 5%椰汁 蔗糖7.5 g/L。文心兰PLBs在5周内的增殖生长曲线呈倒"V"字形,第3周的增殖速度达最高峰,而固体培养基PLBs增殖速度较慢,生长曲线几乎成直线。液体增殖的PLBs分化成苗较固体培养增殖的PLBs差。     

蝴蝶兰原球茎液体增殖培养的研究

以茎尖诱导形成的原球茎为外殖体,采用液体培养方式探讨了不同浓度的激素配比、蔗糖浓度和添加不同量的新鲜液汁对蝴蝶兰原球茎增殖的影响,并比较了液体和固体两种培养方式的原球茎增殖特征.结果表明:不同浓度的外源激素及其配比对蝴蝶兰原球茎增殖影响不明显,但对原球茎生长影响较大,6-BA 2.0 mg/L+Ad 1.0 mg/L+NAA 0.5 mg/L的激素浓度组合比较适合蝴蝶兰原球茎增殖,蔗糖浓度对蝴蝶兰原球茎增殖和生长影响较大,适合蝴蝶兰PLBs增殖的蔗糖浓度为20 g/L,添加10 % ～40 %的新鲜椰汁对蝴蝶兰原球茎增殖生长有明显促进作用;原球茎液体增殖培养的时间不宜超过4周,5周内连续培养增殖曲线呈倒"V"字形;液体增殖培养的原球茎分化成苗的时间相对较晚.     

纳米碳对离体培养条件下几种植物生长及分化的影响

以食用百合、红掌、大花蕙兰及金昌枣的组培苗为材料,研究不同浓度的纳米碳对几种植物外植体生长及分化的影响。结果表明,MS+6-BA 0.8 mg/L+NAA 0.2 mg/L+纳米碳0.10 g/L,促进百合鳞茎增殖,当纳米碳为0.50 g/L时有利于百合苗的生长及生根;MS+6-BA 1.5 mg/L+IBA 0.2 mg/L+纳米碳0.04-0.06 g/L适用于红掌腋芽外植体愈伤组织诱导和分化,诱导率达90%以上,分化率达97%;加入纳米碳1.0-2.0 g/L时能够明显抑制大花蕙兰原球茎继代培养中的褐化现象;MS+6-BA 1.5 mg/L+IBA 0.5 mg/L+纳米碳0.05 g/L,利于金昌枣芽增殖,当纳米碳为0.15 g/L时,芽长势健壮。     

月季‘阿维兰’离体快繁技术

取‘阿维兰'月季带侧芽幼嫩茎段,以改良MS为基本培养基,加入不同浓度的6-BA、NAA及活性炭,通过对不同配方条件下月季诱导分化及生长状况的观察,分析不同生长调节剂浓度以及有无活性炭对其分化及扩繁的影响,并以不同浓度的IBA和NAA进行生根培养.结果表明,'阿维兰'月季离体侧芽诱导的适宜培养基为改良MS 6-BA2.0mg/L NAA0.3mg/L;最适增殖培养基为改良MS 6-BA 2.0mg/L NAA0.1mg/L;最适生根培养基为改良1/2MS NAA0.5mgtL.     

茅苍术胚培养与快速繁殖(简报)

茅苍术种胚在MS 6-BA 1.0mg/L NAA 0.2mg/L培养基上可快速生长并分化出不定芽;不定芽在MS 6-BA 2.0mg/L NAA 0.2mg/L培养基上快速增殖,45d增殖倍数为4.5;增殖芽在1/2 MS NAA 0.5mg/L培养基上快速生根。     

红掌茎段侧芽离体快繁技术研究

以红掌嫩茎为外植体,诱导侧芽萌发,并进行增殖和生根培养,研究不同生长调节剂浓度配比对茎段侧芽萌发、增殖、无菌苗生根的影响以及增殖培养过程中愈伤组织的抑制等因素。结果表明,侧芽诱导的适宜培养基为MS+6-BA 2.0 mg/L+NAA 0.5 mg/L,萌发率达87.5%;最适增殖培养基为MS+6-BA 0.8 mg/L+NAA 0.2 mg/L+VB2 8.0 mg/L,增殖系数3.8;最适生根培养基为1/2MS+NAA 0.5 mg/L,生根率98%;在增殖培养基中添加适量VB2能较好地抑制愈伤组织的生成,防止愈伤组分织分化形成芽,从而达到以芽繁芽的目的。     

植物生长调节剂对花叶开唇兰原球茎增殖和分化的影响

应用正交设计法探讨3种植物生长调节剂对花叶开唇兰原球茎增殖和分化的影响。结果表明,6-BA对原球茎增殖和分化作用显著,NAA和KT作用不显著;原球茎增殖最佳培养基为改良KC+6-BA 2.0 mg/L+NAA 1.5 mg/L+KT 0.2 mg/L,原球茎分化最佳培养基为改良KC+6-BA 1.0 mg/L+NAA 0.5 mg/L。     

西蒙得木茎段组织培养中腋芽生长和增殖的激素调节

6-苄基嘌呤(6-BA);赤霉素(GA_3)和吲哚乙酸(IAA)对西蒙得木茎段的腋芽生长和增殖都可起促进作用,三种激素间有复杂的相互关系。6-BA的作用尤为明显和重要,适宜浓度为1—2mg/L。GA_3浓度为1mg/L时对腋芽生长和增殖有促进进作用,但浓度再增加,长出的腋芽数减少,且芽条生长不良。6-BA与GA_3组合使用,能明显促进腋芽生长,最好组合为6-BA_1—2mg/L GA_3 0.5mg/L:若6-BA浓度升高时,出芽量虽增多,但芽条生长不良。IAA1mg/L或2mg/L时添加不同浓度的6-BA,长出的腋芽数均比单独使用IAA要多;在加低浓度的6-BA(1—2mg/L)时抽出的腋芽数比单独使用6-BA(1—2mg)的少,但芽条生长健壮;在加入较高温度的6-BA(3—5mg/L)时抽出的腋芽数比单独使用6-BA(3—5mg/L)时多。     

硝酸镧对霍霍巴多芽苗生长的促进作用及植株再生

在增殖培养基(改良MS 2mg／L6-BA 0．5mg／LG3)中添加1～3．5mg／L硝酸镧,对霍霍巴试管苗生长有显著促进作用,2．5mg／L硝酸镧对多芽分化有极显著的促进;在生根培养基(改良1/2MS 3mg／LIBA 1．5mg／LNAA)中,1～2mg／L的La(NO3)3对根的分化有显著的促进作用,生根率提高,最佳浓度为2．0mg／L。过高的浓度对试管苗生长有一定抑制。试验表明,不同器官对硝酸镧的敏感程度不同。取2cm高以上的霍霍巴试管苗,用25mg／L IBA或NAA处理30min,扦插于沙基质中。保持室温20～30℃。50d后揭去覆膜,保持光强3000 lx,相对湿度85％以上。正常管理条件下成活率达75％。     

圆叶蔓绿绒快速繁殖条件的优化

以带侧芽茎段为外植体,对圆叶蔓绿绒组织培养及快速繁殖的研究表明,不同生长调节剂组合对圆叶蔓绿绒的离体形态发育及增殖均有重要影响。其中适合侧芽生长和不定芽分化的组合为BA 3.0mg/L+ IAA 0.5mg/L。采用BA 0.8mg/L+ KT 0.8mg/L+IAA 0.05mg/L的组合可在4周内可获得7.0的增殖系数。培养基中添加适量活性炭对生根有促进作用。适宜量为1.0g/L。     

墨兰的无菌播种和植株再生

墨兰的无菌播种结果发现,在不添加细胞分裂素的培养基上,种子可以发芽,但只有原球茎和根状茎产生;不可能进一步分化成苗,只有在含有不同激素成分的MS或KnudsonC培养基上,才有可能诱导芽的分化,其中以附加6-BA 0.5-1.0mg/L+NAA 0.1mg/L诱导效果最佳,在附加6-BA 2.0mg/L+NAA 0.4mg/L的MS培养基中,能加速芽的增殖,根状茎转入含有相同激素成分的液体增殖培养基中振荡培养,可大大提高芽的分化速率。添加0.5%活性炭对芽的分化有明显增效作用。在附加NAA 0.2mg/L的MS培养基中;幼苗的生根效果最佳。     

6-BA、GA3调控春石斛花芽分化的效应

调查了春石斛假鳞茎封顶和增粗的生长情况,并以不同浓度6-BA、GA₃溶液进行灌根处理,观测其对花芽分化的效应。结果表明,假鳞茎的增粗阶段在9月15日至10月10日,经过约50 d增粗生长,假鳞茎粗度迅速增加。其间,6-BA、6-BA+GA₃处理对春石斛花芽分化有促进作用。3种6-BA浓度作用效果明显,最佳为400 mg/L;6-BA添加低浓度GA₃,对花芽分化有增效作用,其中以6-BA 200 mg/L添加GA₃ 50 mg/L效果最佳。GA₃作用效果不稳定,浓度为25 mg/L时叶片脱落不明显,高于50 mg/L则叶片大量脱落。     

油松体细胞无性系的建立

以油松（Ｐｉｎｕｓｔａｂｕｌａｅｆｏｒｍｉｓ）种胚为外植体,通过器官发生途径建立体细胞无性系。以ＭＳ为基本培养基,在芽的诱导和增殖培养基中,附加植物激素,以６ＢＡ＋ＫＴ和ＮＡＡ效果最好,两者的配比为５－１０：１,６ＢＡ和ＫＴ的浓度分别均不超过１ｍｇ／Ｌ。同时根据培养物的发育状态交替使用活性炭,则对芽的增殖有明显的促进作用。选择发育状态较幼嫩的刚抽茎的外植体,诱导生根,在１／２ＭＳ＋ＫＴ０．１＋ＩＢＡ０．１＋ＮＡＡ０．１ｍｇ／Ｌ的培养基中,生根率达３２．％。体细胞胚胎发生途径建立细胞无性系也已取得成功,筛选出可继代培养的胚性胚柄团无性系,在胚轴和子叶上诱导出成熟的体细胞胚并获得完整小植株。     

胡杨遗传转化体系的建立及抗生素浓度的优化

探讨了不同激素浓度对胡杨叶片分化以及卡那霉素、G418、羧苄青霉素和头孢霉素4种抗生素对胡杨不同培养阶段外植体生长、分化或生根的影响,确定了由农杆菌介导的胡杨遗传转化研究中抗生素种类和转化体的筛选浓度,建立了适于胡杨叶片转化的遗传转化体系。结果表明:胡杨叶片再生的最佳培养基为MS+BA0.5mg/L+NAA0.1～0.2mg/L+白砂糖25g/L+琼脂5g/L;在叶片转化筛选阶段,卡那霉素和G418的适宜浓度分别为10mg/L和7.5mg/L,羧苄青霉素和头孢霉素的适宜浓度为200～600mg/L和200～400mg/L;在抗性芽生根培养时,卡那霉素和G418分别为15～20mg/L和10～15mg/L,羧苄青霉素和头孢霉素为200～800mg/L和200～600mg/L。     

向日葵离体再生体系的建立

为了建立高效的向日葵离体再生体系,从基因差异、外植体取材、生长素和细胞分裂素浓度、附加物的添加等方面出发,对向日葵愈伤诱导、分化、生根等过程进行了系统优化。结果表明：杂交材料相对于自交材料更容易实现再生;最佳外植体是生长4 d的子叶;最佳愈伤诱导培养基是MS培养基 (MS) +2.0 mg/L 6-苄基腺嘌呤 (6-BA)+0.5 mg/L奈乙酸 (NAA)+1.0 mg/L激动素 (KT),诱导率最高可达100％;最佳分化培养基是MS+0.2 mg/L 6-BA+0.5 mg/L NAA+0.3 mg/L KT+0.3 mg/L硝酸银 (AgNO3)+0.2 g/L活性炭 (AC),芽分化率可达71％;最佳生根培养基是1/2 MS+0.6 mg/L吲哆丁酸 (IBA),生根率最高为77％。方差分析表明,材料基因型、外植体生长时间、激素、AgNO3、AC对向日葵再生呈现显著性影响。     

The Exogenous Hormornal Control of the Development of Regenerated Flower Buds in Hyacinthus orientalis

The critical role of exogenous hormone on inducing the initiation of different floral organs in the regenerated flower bud and controlling their numbers was further evidenced. The initiation of the flower buds was first induced from the perianth explants of Hyacinthus orientalis L. cv. White pearl by a combination of 2 mg/L 6-BA and 0.1 mg/L 2,4-D, and then a continuous initiation of over 100 tepals (a flower bud of H. orientalis in situ has only 6 tepals) was successfully controlled by maintenance of such a hormone concentration. However, a change of hormonal concentration (2 mg/L 6-BA and 0-0.000 1 mg/L 2,4-D) caused cessation of continuous initiation of the tepals but gave rise to induction of stamen initiation. Keeping the changed hormone concentrations could successfully control the continuous initiation of over 20 stamens (a flower bud of H. orientalis in situ has only 6 stamens). The experiment showed that the number of identical floral organs in the regenerated flower buds can be controlled by certain defined concentrations of the exogenous hormones, and the amount of the induced identical floral organs has no effect on the differentiation sequence of the different floral organs in the regenerated flower bud. Based on a systematic research on controlling the differentiation of the floral organs from both the perianth explants and the regenerated flower buds by the exogenous hormones in H. orientalis over the past decade, the authors put forward here a new idea on the role of phytohormone in controlling the automatic and sequential differentiation of the different floral organs in flower development. The main points are as follows: 1. the development of flower bud in plant is a process in which all of the floral organs are automatically and sequentially differentiated from the flower meristem. 2. Experiments in vitro showed that the effect of exogenous hormones in controlling the initiation of different floral organs is strictly concentration dependent, i.e., one kind of the floral organ can continuously and repeatedly initiate from the flower meristem as long as it is maintained in that specific concentration of the exogenous hormone which is suitable for the initiation of that particular kind of floral organ. 3. It shows that the flower buds in situ must be automatically able to adjust the endogenous hormonal concentrations just after the completion of the differentiation of one whorl of floral organ to suit the differentiation of the next whorl. Thus, the phytohormone in different concentrations takes after many change-over switches of the organ differentiation and plays a connective and regulatory role between the differentiation of every two whorls of the floral organ. In other words, these change-over switches play the roles of inhibiting the expression of the genes which control the initiation of the floral organs in the first whorl, meanwhile, activating the expression of the genes which control the initiation of the floral organs in the second whorl during the successive initiation of the different floral organs from the flower bud. It results in the automatic and sequential initiation of the various floral organs from the floral meristem.      

沙冬青愈伤组织对培养基的特殊要求和球形胚状分化结构的诱导

通过对沙冬青未成熟子叶进行离体培养,获得了愈伤组织。基本培养基采用Ｂ５,再配合ＭＳ培养基的铁盐成分,对沙冬青未成熟子叶愈伤组织的诱导和生长效果最佳。愈伤组织的诱导和生长对培养基的附加成分要求十分严格,２,４－Ｄ浓度为０．５ｍｇ／Ｌ,６－ＢＡ浓度为０．５ｍｇ／Ｌ,蔗粮不宜超过２％。愈伤组织最初为乳白色透明松散状态,生长比较缓慢,极易褐化死亡。如果培养基中的蔗糖浓度低于２％,两周后未成熟子叶愈伤组织逐     

Plant Regeneration from Cell Supension Derived Protoplasts of Heracleum moellendorffii

Heracleum moellendorffiz Hance is a herb belonging to Umbelliferae used in traditional medicine in China. The young stem-nodes were induced for callus formation on MS medium containing 1 mg/L 2,4-D. After subcultured for about five months, the embryogenic calli were used for cell suspension culture. The protoplasts were prepared from this suspension by digestion with enzyme mixture containing 1. 5% cellulase Onozuka R-10 +0. 3% macerozyme R-10 + 0. 5% snailase + 5 mmol CaCl2 + 0. 6 mol/L mannitol, at pH 5.8, and cultured in modified MS and modified N6 media with 0.3 % agarose. They divided after 3 days and developed into small cell colonies after about 2 weeks. From this time on, the glucose concentration in the culture media was decreased to 0. 2 mol/L,which led to futher growth of the colonies to small calf . After a period of proliferation on solid medium with 0. 5 mg/L 2,4-D, the calli were transferred to a medium with 0. 1 mg/L zeatin on which somatic embryos differentiated and developed to plantlets