滇牡丹愈伤组织继代培养中的褐化防治

以滇牡丹Paeonia delavayi Franch成熟种胚诱导出的愈伤组织为试验材料,研究了PVP、VC、结冷胶、AgNO3、水解乳蛋白(LH)5种抗褐化剂在牡丹愈伤组织继代培养过程中的褐化控制作用和对愈伤组织生长量的影响。结果表明:愈伤组织褐化主要发生在第一次继代转接后的10 d内;在愈伤组织两次继代培养过程中,3.5g/L的结冷胶处理抗褐化效果最好,LH和AgNO3在整个继代过程中也表现了一定的抗褐化性,PVP和VC抗褐化性能相对较差。AgNO3对愈伤组织生长具有明显的促进作用。     

大花重瓣非洲紫罗兰植株再生过程中内源激素变化规律的研究

以大花重瓣非洲紫罗兰为外植体,探讨了在植株再生过程中,内源激素的变化规律.结果表明:植株再生过程中不同阶段的内源激素的变化规律不同.适当添加IAA有利于愈伤组织的形成;ABA在愈伤组织形成过程中含量下降,在继代和分化过程中一直保持较低水平,是此过程中的负调控因子.GA在继代和分化过程中均呈现上升趋势,说明其对愈伤组织的大量形成和分化有重要作用.     

牡丹愈伤组织诱导和继代培养体系的建立

以牡丹为材料,研究了不同外植体、激素对愈伤组织诱导和继代增殖的影响,结果表明:①牡丹子叶、胚轴是诱导愈伤组织的最佳外植体;②牡丹子叶愈伤组织诱导的最佳培养基为:MS+Ca2+;③牡丹愈伤组织增殖的最佳培养基为:MS+2,4-D 0.5 mg.L-1+NAA 1.5 mg.L-1+Vc 500 mg.L-1;④在培养基中添加500 mg.L-1Vc或去除Cu2+,具有较好的抗褐化效果。     

雷公藤胚性愈伤组织再生植株的增殖及其稳定性

以雷公藤胚性愈伤组织为材料,研究继代培养时间对植株再生和基本培养基、生长素浓度及其与细胞分裂素组合对再生苗增殖的影响.结果表明:在继代培养过程中,愈伤组织再生率和再生苗数经历了一个由低到高再到低的过程,以8个月到1年之间的胚性愈伤组织的再生能力最强,达100%,每块愈伤组织平均形成的芽数也比较多,雷公藤胚性愈伤组织在继代20个月后已经丧失再分化能力.愈伤组织再分化过程易发生变异,从叶形上看,再生苗出现6种类型的变化.再生苗茎段增殖以1/2MS为基本培养基附加2.0 mg·L-1IBA为好,以1/2MS+2.0mg·L-1IBA+O.5 g·L-1活性炭的培养基有利于芽苗生根培养,生根率达100%.移栽至珍珠岩与腐殖质土(1:1)的混合基质中成活率达96%.     

胡杨愈伤组织继代增殖和器官发生中蛋白分子标记的研究

利用十二烷基磺酸钠聚丙烯酰胺凝胶电泳 (SDS PAGE)和等电聚焦双向电泳 (IEF 2D)对胡杨不同类型愈伤组织和愈伤组织分化不定芽过程中表达的蛋白进行了研究。发现 :不同类型愈伤组织中表达的蛋白存在着明显的差异。在光照和培养基中BA NAA值为 1时诱导产生的有极强器官发生能力的茎基愈伤组织 ,其蛋白组分明显地少于经过继代培养、器官发生能力明显下降的愈伤组织 ,表明了茎基愈伤组织本身分化程度低。实验获得了不同类型愈伤组织和愈伤组织分化不定芽过程中不同阶段所表达的特异蛋白 ,表现为经过黑暗和培养基中BA NAA值为 0 5的继代增殖培养 ,愈伤组织产生了特异的 2 4 5kDa和 5 8 6kDa的标记蛋白带 ,并且也表达了其器官发生时表达的 19kDa和 31kDa蛋白带。茎基愈伤组织在光照和BA NAA值为 0 5的条件下进行器官发生诱导并且随着愈伤组织形成分生细胞团块和不定芽原基 ,明显地表达了 2 0kDa和 5 5kDa蛋白带 ,在 2 0kDa蛋白带中含有pI为 5 5～ 6 5的特异蛋白。pI为 6 5～ 7 5的 4 3kDa为不定芽发生前期表达的特异蛋白。文中就愈伤组织器官发生能力与其表达蛋白的关系进行了讨论。     

白桦愈伤组织染色体制片方法及数目变异研究

该文探讨白桦愈伤组织染色体的制片方法,并对白桦愈伤组织及其再生植株的染色体数目变异进行了研究。结果表明：取继代培养1周的白桦愈伤组织,用0.2%的秋水仙素溶液预处理1.5～2 h,1 mol/L HCl室温（18～20℃）解离10～15 min或2%纤维素酶和果胶酶混合液室温解离30 min,压片后易获得分散、染色效果好的细胞分裂中期染色体图像。随着继代次数增加,白桦愈伤组织染色体数目变异的频率及范围增大。在愈伤组织分化过程中,二倍体细胞占有优势,能够分化为正常的植株,愈伤组织再生植株的变异频率低于愈伤组织。     

香蕉薄切片愈伤组织再生体系的建立研究

以巴西香蕉品种的薄切片为外植体高频地诱导出愈伤组织,并诱导出了不定芽,获得了再生植株。在实验过程中,利用正交实验中筛选出愈伤组织诱导的最佳培养基为MS+2,4-D 4.0 mg.L-1+KT 1.0 mg.L-1+NAA 1.0 mg.L-1+活性炭200 mg.L-1,诱导率可达100%,愈伤组织增殖的最佳培养基是MS+2,4-D 2.0 mg.L-1+6-BA 0.5 mg.L-1+NAA 0.5 mg.L-1+活性炭100 mg.L-1,继代3-4次的愈伤组织在6-BA浓度为5.0 mg.L-1的分化培养基上能分化出不定芽,继代10次之后愈伤组织在继代培养基上开始出现褐化现象,愈伤组织增殖明显受到抑制,褐化的愈伤组织逐渐死亡。     

黑荆树愈伤组织诱导、增殖与分化

以黑荆树叶片为外植体,MS培养基为基本培养基,研究了外源激素对愈伤组织诱导、增殖及分化的影响。结果表明:外源激素6-BA与IBA组合、6-BA与2,4-D组合、TDZ与IBA组合均可诱导愈伤组织,但诱导率差异达极显著水平;不同外源激素组合上诱导的愈伤组织,其增长率为165.21%~208.53%,其中以6-BA与IBA组合的增长率最大;在添加TDZ与IBA的培养基上易分化出不定芽,但不同培养基上诱导的愈伤组织对不定芽的分化率和芽的生长影响达极显著差异。综合考虑,筛选适宜愈伤组织诱导、增殖培养基为MS+1.5 mg/L 6-BA+0.5mg/L IBA,诱导率为43.36%,平均增量和增长率分别为1.04 g和208.53%;最适宜继代时间为18~21 d;最适宜愈伤组织分化培养基为MS+0.05 mg/L TDZ+0.2 mg/L IBA,分化率为51.85%,每块愈伤组织上分化5~8个芽点。     

毛竹愈伤组织培养中褐变现象的研究

在毛竹组织培养过程中，愈伤组织褐变非常严重，直接影响愈伤组织的继代及分化。通过对激素种类、温度、光照及褐变抑制剂等影响毛竹愈伤组织褐变因素的研究，找到最大限度避免或减轻毛竹愈伤组织褐变的方法。     

枣树茎段试管组培的研究

比较了5种培养基对枣树茎段愈伤组织的诱导和植株再生的效果;3种外植体的诱导分化效果及不同取材时间的诱导效果;表明枣树茎段通过愈伤组织再生植株并进行继代培养是可行的。筛选出了最佳诱导分化培养基及最佳取材时间和适宜的外植体,得到了理想的试管苗,并进行了生根试验,取得了满意的结果。     

Effect of organogenesis on steroidal saponin biosynthesis in calli cultures of Ruscus aculeatus

Calli cultures derived from Ruscus aculeatus rhizomes were investigated for their potential to biosynthesize saponins. The capacity of undifferentiated tissues to form steroidal saponins is very limited, but when the calli developed organogenesis, mainly aerial shoots and roots, the saponin production increased significantly. Plantlets regenerated from aerial shoots of Ruscus calli showed a saponin pattern similar to that of the callus cultures but the levels of saponins found in the aerial part and roots were significantly greater.     

落叶松不同类型胚性和非胚性愈伤落叶松不同类型胚性和非胚性愈伤

以华北落叶松继代、增殖培养的不同类型的胚性及非胚性愈伤组织为材料，测定其氨基酸、糖类、酚酸、激素以及离子的含量，结果表明氨基酸含量的差异十分明显，质地硬的非胚性愈伤组织中18种游离氨基酸含量为胚性愈伤组织的2倍以上，经ABA处理后，胚性愈伤组织中多糖及邻苯二酚含量急剧升高；三糖含量与不同基因型密切相关；乙烯释放量在非胚性愈伤组织中高于胚性愈伤组织；金属离子CO     

Selection, regeneration and protein profile characteristics of NaCl-tolerant callus ofRobinia pseudoacacia L.

Changes in callus growth, differentiation potency and protein profile induced by salt stress were investigated in the calli induced from a hypocotyl ofRobinia pseudoacacia L. The NaCl treatment evidently obstructed the callus growth and the influences seemed to continue to be present in the selected calli even after they were restored in NaCl-free medium that could partially recover the callus growth, and some polypeptides were found to restore. The calli selected and restored intermittently every two weeks for one year completely lost their differentiation potency. The NaCl-tolerance test of the regenerated plantlets revealed that part of the plantlets that directly regenerated from the selected NaCl-tolerant calli were able to survive in 0.2 M NaCl, while most of the plantlets regenerated from the NaCl-nonselected calli or from restored calli were obviously damaged by this treatment. The SDS-PAGE assay indicated that a 54 kDa polypeptide increased distinctly to a very high level while a 41 kDa polypeptide decreased to a lower level in almost all the NaCl-tolerant plantlets. This observation led to the suggestion that the changes may play an adaptive role in allowing plantlets to survive under saline conditions.     

不同盐浓度对木麻黄无性系愈伤组织的影响

以短枝木麻黄4个无性系幼枝茎段为外植体，在不同NaCl浓度的培养基上进行愈伤组织诱导，观察不同盐浓度对木麻黄外植体愈伤组织分化的影响。试验结果表明：培养基中NaCl浓度达到0．064mol／L(0．374％)，比正常浓度高出320倍的情况下木麻黄外植体表现出很强的耐盐性，仍然能诱导出正常的愈伤组织；不同的NaCl浓度的各个处理之间愈伤组织的诱导率、大小、出芽率、继代愈伤组织的大小均没有显著差异；但无性系之间的愈伤组织的诱导率及愈伤组织的大小均有极显著差异；不同继代间愈伤组织的大小有极显著差异。     

Effect ofl-phenylalanine on rooting of shoots from axillary buds of adultLarix leptolepis

Rooting of shoots derived from axillary buds was examined to establish an efficient shoot culture system of clonal micropropagation in adult tree ofLarix leptolepis Gord. (Japanese larch). Nine out of ten shoots induced calli (90%) on their shoot bases, and the two of them formed root primordia with a red pigment (20%) on the calli surface within 5 weeks after culturing on modified Murashige and Skoog (MS) medium supplemented with 1.5 μM of indolebutyric acid (IBA) and 1.5 μM of naphthaleneacetic acid (NAA). However, the primordia did not elongate actively. The addition of 10 mMl-phenylalanine in the MS medium with the auxins resulted in the formation of roots at high frequency, about 80%, and they elongated actively. Although callus was formed in all the shoots cultured on the medium withl-phenylalanine, it appeared that the callus development was less as compared to the medium withoutl-phenylalanine. Consequently, the rooting might be associated with the suppression of the induced callus.     

Mevalonate kinase activity during different stages of plant regeneration from nodular callus cultures in white pine (Pinus strobus)

Mevalonate kinase (MK) catalyzes a step in the isoprenoid biosynthetic pathway, which leads to a huge number of compounds that play important roles in plant growth and development. Here, we report on changes in MK activity in white pine (Pinus strobus L.) during plant regeneration by adventitious shoot organogenesis from cotyledons of mature embryos, including nodular callus induction, shoot formation and rooting. Nodular calli were induced from Pinus strobus (PS) embryos by culture in nodular callus induction medium in a 0-, 8- or 16-h photoperiod. Mevalonate kinase activity peaked in nodular calli after three weeks of culture on nodular callus induction medium in a 16-h photoperiod, whereas frequency of nodular callus formation peaked after 4 weeks of culture on nodular callus induction medium in darkness. During adventitious shoot formation, MK activity peaked in shoots derived from dark-grown nodular calli after 3 weeks on bud formation medium, and frequency of shoot formation was highest in dark-grown nodular calli cultured on bud formation medium for 4 weeks. During rooting, MK activity peaked 2 weeks after transfer of adventitious shoots to rooting medium and rooting frequency was highest in adventitious shoots after 3 weeks on rooting medium. Although during nodular callus induction in darkness MK activity was inversely related to frequency of nodular callus formation, MK activity was highly correlated with frequency of shoot formation and with rooting frequency. The observed increase in MK activity preceding rooting suggests that MK could serve as a marker for rooting of white pine shoots in vitro.     

抗生素对火炬松的器官发生植株再生的影响(英文)

本文研究了氨苄青霉素、羧苄青霉素和头孢霉素等3种抗生素对火炬松愈伤组织的生长和分化及不定芽生根的影响。结果表明,头孢霉素最有利于愈伤组织的诱导和生长,羧苄青霉素最有利于芽的分化,氨苄青霉素降低了不定芽的生根频率。所有试验的3种抗生素提高愈伤组织的形成和芽再生,但降低了芽的生根频率。这些结果表明,选择合适的抗生素对优化火炬松遗传转化体系有重要作用。图3表4参25。     

盾叶薯蓣愈伤组织培养及其高产系的筛选

以盾叶薯蓣的块茎、茎、叶和茎尖为材料,对其进行了愈伤组织培养,并根据愈伤组织个体间生长速度和皂甙含量差异,进行了高产系的筛选．结果表明：盾叶薯蓣不同外植体（块茎、茎、叶、茎尖）均能诱导出愈伤组织,其中以茎尖形成愈伤组织最快,皂甙元含量最高;不同培养基、ｐＨ 值、接种量、温度、激素等因子对愈伤组织的形成、生长及皂甙元含量有很大的影响;改良 Ｍ Ｓ培养基、２,４－Ｄ 浓度为２．０～４．０ ｍ ｇ／Ｌ、ＢＡ 为 ０．５ ｍ ｇ／Ｌ 的激素配比使愈伤组织的生长量、产物含量最高;对无性系进行筛选和 ＨＰＬＣ 检测,以 １２号无性系生长速率和皂甙元含量最高,其皂甙含量达 ３．７％ ,生长量比普通系高２１．５％ ．     

野葛愈伤组织诱导与不定芽分化

采用MS为基本培养基，附加NAA、6-BA和IAA3种激素．诱导野葛不同外植体愈伤组织形成；再将愈伤组织接种在不同浓度6-BA的MS培养基上．分别在光照和黑暗条件下进行不定芽的诱导。结果表明．野葛幼叶、茎段和顶芽在适宜激素的诱导下均能形成愈伤组织，但不同激素组合其愈伤组织诱导率不同。幼叶愈伤组织诱导的最适培养基为MS NAA1．0mg／L 6-BA1．0mg/L．其诱导率为75％．茎段愈伤组织诱导的最适培养基为MS NAA1．0mg／L 6-BA3．0mg／L IAA0．2mg/L，其诱导率为70％。不同外植体形成的愈伤组织其不定芽的分化诱导率不同．茎段愈伤组织在不同浓度6-BA下均能诱导出不定芽；光照有利于芽的分化．在光培养条件下，茎段愈伤组织不定芽平均诱导率为66．7％。