外源激素在诱导风信子花被外植体不同部位细胞再生花芽中的作用

外源激素诱导风信子（Ｈｙａｃｉｎｔｈｕｓ ｏｒｉｅｎｔａｌｉｓＬ．）同一发育时期花被外植体不同部位细胞再生花芽的实验表明∶１． 诱导花被外植体细胞再生花芽,外源激素是必需的;２． 仅有细胞分裂素就可以诱导花芽再生,生长素并不是必需的;３． 花被外植体上的不同部位的细胞再生花芽时,需要不同浓度的外源激素． 单独加６－ＢＡＰ或玉米素２ ｍ ｇ／Ｌ可以诱导花被下部的细胞再生花芽;６－ＢＡＰ或玉米素２ ｍ ｇ／Ｌ和２,４－Ｄ ０．１ ｍ ｇ／Ｌ的组合有利于花被中部的细胞再生花芽;６－ＢＡＰ或玉米素２ ｍ ｇ／Ｌ和２,４－Ｄ １．０ ｍ ｇ／Ｌ的组合能促进花被上部的细胞分化花芽     

小麦外稃和内稃愈伤组织直接再生小穗和雌蕊状结构的研究

以小麦稃片为外植体,在MS附加不同外源激素的培养基上,成功地从愈伤组织上直接诱导了小穗和雌蕊状结构的再生。实验表明,附加2,4-D 2mg/L和6-BAP 0.1mg/L有利于外植体形成愈伤组织;附加2,4-D 1.0mg/L和6-BAP 2mg/L有利于外稃和内稃愈伤组织直接再生小穗;附加2,4-D 0.01mg/L和6-BAP 1.0mg/L有利于内稃愈伤组织直接再生雌蕊状结构。本文还观察了小穗和雌蕊状结构再生过程的形态学变化,并对诱导花芽和性器官再生的某些要点进行了讨论。     

In Vitro Induction of Regeneration of Fruit-Like Structure in Lycopersicon esculentum Mill.

It is possible to induce in vitro regeneration of fruit-like structures from tissue pieces excised from young pistil and young fruit of Lycopersicon esculenturn Mill. These fruit-like structures could be cultured in vitro to maturation, meanwhile they became red coloured. Under dissection it was observed that the fruit-like structure consists of fruit flesh and fruit coats without seeds and plancentae. Tests of exogenous hormones and explant ages revealed: 1. Supplement with exogenous cytokinins alone could induce the explants excised from the pistils to regenerate the fruit-like structures, and the highest induced frequency (50%) was obtained when 0. 5 mg/L zeatin was supplemented. Exogenous auxin seems unnecessary. 2. When tissues of young fruit were used as explants, all explants excised from 4—12 mm (diameter) young fruits could be induced to differentiate the fruit-like structures on medium supplemented with exogenous hormones. However, the hightest frequency of induction (62.5%) was obtained only in tissues of explants excised from 8 mm (diameter) sized fruit and cultured on medium supplemented with 2 mg/L of 6-BAP and 1 mg/L of NAA where regermination of the fruit-like structures took place. In order to investigate the expression of cell totipotency arisen in the regeneration of the fruit-like structure a concept of partial expression of plant cell totipotency was proposed and discussed.     

Study on in Vitro Regeneration of Style-Stigma-Like Structure in Crocus sativus L.

Effects of age and variety of explant as well as of exogenous hormones on regeneration of style-stigma-like structure in vitro were observed in saffron. (1) Explants excised from stalk, perianth, anther, ovary, style and stigma showed different reactions in vitro, only young perianth and style could regenerate the style-stigma-like structures; (2) Age of the perianth explant had obvious effect on induction frequency, and the explants excised from an inflorescence of 24mm in length showed the highest frequency (37.5%) and (3) Experiments on exogenous hormones made clear that supplement with K 5mg/L and NAA 4mg/L was advantageous to regeneration of style-stigma-like structure. Induction frequency reached 35.3%. Supplement with 6-BAP 7mg/L and NAA 9mg/L could promote the formation of stigma like structures.     

离体培养下番红花花柱—柱头状物再生的研究

番红花(Crocus sativus L.)幼花被和幼花柱可以诱导花柱-柱头状物的再生,长度为24mm 的花芽上分离的花被具有最高的诱导频率(37.5%);花被外植体附加 K5 mg/L 和NAA 4mg/L 有利于再生花柱-柱头状物,频率达到35.3%;附加6-BAP 7 mg/L 和 NAA9mg/L 则有利于柱头状物的大量形成。     

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.      

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。     

盾叶薯蓣离体成花的影响因素及组织学观察

研究了居群、外植体类型、激素、铁盐对盾叶薯蓣（Dioscorea zingberensis C．H．Wright）离体成花的影响,建立了从花序切段高效直接再生花序和小花的实验体系。以不同的培养基对5个居群盾叶薯蓣的花序进行离体培养,利用石蜡切片技术观察再生花序的发生。结果表明5个居群的盾叶薯蓣都能直接再生花序,与花器官相关组织的外植体均具有不同程度的花序再生能力。培养基的组成对离体花序诱导率有很大影响,BA促进形成花序,KT与高浓度的铁盐促进形成营养芽。其中,MS＋2．0mg／LBA＋0．2mg／LNAA最有利于花序再生及发育。切片观察表明,离体再生的花序主要发生于花序轴与花蕾交界处的苞片以及小花的花被片表层细胞。     

A Preliminary Study on Direct Regeneration of Flower Buds from Peduncle Calli in Dracaena fragrans cv. massangeana

Flower buds were directly regenerated from calli in vitro in the woody plant Dracaena fragrans cv. massangeana Hort. On modified MS medium supplemented with 1.0 mg/L 6-BA and 1.0 mg/L IBA, two kinds of calli, A and B, were formed from the peduncle explants cultured for 5 months. Calli A were loose and on their surface there were many irregular granule-like structures (GLC); Calli B were compact and had bigger tumor-like structures (TLC) on their surface. When the GLC and TLC were transferred onto the medium respectively with 0.4 mg/L 6-BA and 1.0 mg/L IBA, flower buds were differentiated directly from the GLC but only vegetative buds and roots were differentiated from the TLC after culturing for 4 weeks. The GLC could be partly transformed into TLC in the continuous passage culture. Assays on hormones revealed that at a fixed IBA concentration of 0.4 mg/L the defferentiation frequency of flower budding was increased as the 6-BA concentration was decreased from 2.0 mg/L to 10 mg/L. Alternatively, at a fixed 6-BA concentration of 2.0 mg/L, the flower budding frequency was increased when the IBA concentration was changed from 0.4 mg/L to 1.0 mg/L. Moreover, the addition of 2.0 mg/L zeatin to the culture medium containing 2.0 mg/L 6-BA and 0.4 mg/L IBA was favorable to the regeneration of the flower buds. Nevertheless supplementing 1.0 mg/L GA3 into the medium on which the calli had differentiated into flower buds, the flower buds would gradually wither after 2 weeks in culture.     

花叶千年木花序梗愈伤组织直接再生花芽的初步研究

在离体条件下,诱导愈伤组织或外植体直接再生花芽已经在许多草本植物上取得成功［１～７］,而就木本植物来说,迄今尚未见到成功的报导。诱导愈伤组织或外植体直接再生花芽所形成的离体培养实验系统将十分有利于研究雌、雄性器官分化和发育所必需的条件［８］和找到所需...     

Further studies on the effects of different auxins and cytokinins on flower formation in thin cell layers of Nicotiana tabacum: The effects of zeatin riboside, dihydrozeatin and dihydrozeatin riboside

Organogenesis in thin cell layers of Nicotiana tabacum L. was studied in relation to the effects of natural and synthetic auxins in combination with various cytokinins. All cytokinins tested, benzyladenine (BA), kinetin, zeatin (Z), zeatin riboside (ZR), N⁶-Δ²-isopentenyl) adenine (IPA), dihydrozeatin [(diH)Z] and dihydrozeatin riboside [(diH)ZR], seem to be active in flower bud formation. In addition to the initiation of flower buds, vegetative buds or roots were also formed on the explants in the presence of BA, Z or IPA as exogenous cytokinins. Only dihydrozeatin and its riboside stimulated the initation of flower buds alone (as is known for kinetin), especially if supplemented with indole-3-acetic acid (IAA) as exogenous auxin. A high number of explants with flower buds was also found with high cytokinin/2,4-D ratios. In these conditions the presence of (diH)Z yielded the higest number of flower buds per explant.     

裂叶悬钩子组织培养研究 Ⅰ.裂叶悬钩子叶外植体培养直接再生不定芽及植物激素对它的影响

本文首次报道裂叶悬钩子(Rubus laciniatus Wild)叶外植体培养在改良的NN~(69)培养基上附加2—4mg/1 6-BA和0.1mg/1 NAA或1—3mg/1 2,4-D和0.1mg/1 NAA,两者都可直接从完整叶片、叶片下切段或叶柄诱导出不定芽。诱导频率达20—48%。而不定芽绝大部分发生在叶轴处或叶柄基部。完整叶片的不定芽诱导率与叶片下切段无差别,但比叶柄基部诱导率要高。6-BA对叶轴处不定芽诱导率比2,4-D的要高。此外,不需继代培养,不定芽数可达10—20个,继代培养一个月左右,每个不定芽能形成丛生芽数可达40一60个。另外,本文还讨论了细胞分化过程中的极性现象。     

梨枣叶片和茎段再生体系的建立

用不同浓度配比的生长素和细胞分裂素诱导梨枣叶片和茎段愈伤组织的产生,并研究了不定芽诱导的最佳配方,建立了梨枣叶片和茎段的再生体系.结果表明,梨枣叶片愈伤组织诱导的最佳培养基为MS+2,4-D 1.5 mg·L-1+6-BA 0.5 mg·L-1;茎段为MS+2,4-D 1.0 mg·L-1+6-BA 0.5 mg·L-1.叶片不定芽诱导的最佳培养基为MS+IBA 0.1 mg·L-1+6-BA 1.5 mg·L-1. AgNO3能阻止叶片外植体褐化并有效地促进叶片愈伤组织分化.茎段能在同一培养基上产生愈伤组织并直接分化出不定芽.     

裂叶悬钩子组织培养研究——Ⅰ.裂叶悬钩子叶外植体培养直接再生不定芽及植物激素对它的影响

本文首次报道裂叶悬钩子(Rubus laciniatus Wild)叶外植体培养在改良的NN⁶⁹培养基上附加2—4mg/1 6-BA和0.1mg/1 NAA或1—3mg/1 2,4-D和0.1mg/1 NAA,两者都可直接从完整叶片、叶片下切段或叶柄诱导出不定芽。诱导频率达20—48%。而不定芽绝大部分发生在叶轴处或叶柄基部。完整叶片的不定芽诱导率与叶片下切段无差别,但比叶柄基部诱导率要高。6-BA对叶轴处不定芽诱导率比2,4-D的要高。此外,不需继代培养,不定芽数可达10—20个,继代培养一个月左右,每个不定芽能形成丛生芽数可达40一60个。另外,本文还讨论了细胞分化过程中的极性现象。     

牛蒡组织培养和高频植株再生的研究

通过对牛蒡(A rctium lapp a L.)不同外植体、不同激素配比的比较研究,建立了牛蒡离体培养高效植株再生体系.牛蒡子叶与下胚轴切段在含2.0 m g/L 2,4-D和0.5~2.0 m g/L BA的M S培养基中愈伤组织诱导率可以达到87%~100%;在1.0~3.0 m g/L NAA和0.5~2.0 m g/L BA的M S培养基上通过愈伤组织间接分化或外植体直接分化形成不定芽,其中愈伤组织分化率可达100%;下胚轴的分化率明显高于子叶,在1.0 m g/L NAA和1.0 m g/L BA的M S培养基上下胚轴直接分化率达77.3%.组织学观察发现牛蒡再生有器官发生和体细胞胚发生两种途径.将生长状态良好的不定芽转至含1.0 m g/L IBA和1.0 m g/L NAA的1/2 M S培养基上生根,移栽,成活率达到93.3%.从诱导愈伤组织到组培苗在珍珠岩中过渡成活,大约需要13周.组培苗次年开花并结实,生长形态特征正常.     

安徽羽叶报春愈伤组织的诱导和培养

为了寻找大批量快速繁殖安徽羽叶报春(Primula merrilliana)的有效办法,笔者以其叶片、叶轴为外植体,用不同浓度生长调节物质进行愈伤组织诱导实验.结果表明:(1)愈伤组织诱导的最适培养基为:MS 2.0mg/L 6 BA 0.5mg/L～1.0mg/L 2,4-D,叶片、叶柄出愈率均达100%.(2)MS培养基加0.1mg/L 2,4～D和1.0mg/L～1.5mg/L TDZ对叶柄的诱导效果较佳,其诱导率达81%,而且在这种生长调节物质浓度下的愈伤组织可直接分化出不定芽.     

Regeneration of tepals,stamens and ovules in explants from perianth of Hyacinthus orientalis L. importance of explant age and exogenous hormones

Regeneration of tepals, stamens and ovules from perianth explants of Hyacinthus orientalis L. in different developmental stages could be controlled by means of exogenous hormones. Perianth explants in a relatively early stage of development were competent for differentiation of tepals on Murashige and Skoog (MS) medium supplemented with 2 mg·1^-1 N⁶-benzylaminopurine (BAP) or zeatin and 0.1 mg·1^-1 2,4-dichlorophenoxyacetic acid (2,4-D). Perianth explants in a later stage of development regenerated stamens and ovules, and marked difference was observed in the activity of BAP and zeatin in this regard. Zeatin stimulated more strongly stamen formation, while BAP enhanced ovule formation. Thus, stamens were formed when the explants were cultured for four months on medium with 2 mg·1^-1 BAP and 0.1 mg·1^-1 2,4-D and then transferred to medium with 0.2 mg·1^-1 zeatin and 0.005 mg·1^-1 1-naphthaleneacetic acid. On the other hand, differentiation of ovules occured in explants cultured for two weeks on the former medium and then transferred to medium with 0.1 mg·1^-1 BAP and 0.01 mg·1^-1 2,4-D. Although ovule formation could also be obtained with 2 mg·1^-1 BAP alone, it was substantially enhanced by the presence of 0.1 mg·1^-1 2,4-D in the medium in the early stages of culture. The results demonstrate the importance of both the developmental stage of the source organ from which explants are excised and of the hormone composition of the medium for the regeneration of different floral organs by perianth explants of Hyacinthus.Abbreviations BAP N⁶-benzylaminopurine - 2,4-D dichlorophenoxyacetic acid - NAA 1-naphthaleneacetic acid     

软枣猕猴桃试管苗叶片和茎段的愈伤组织诱导及植株再生

从软枣猕猴桃试管苗茎段和叶片诱导出愈伤组织并得到再生植株。茎段外植体容易愈伤化,但其愈伤组织难以分化,叶片外植体不易愈伤化,但其愈伤组织容易分化。ＭＳ培养基分别附加ＢＡＰ、Ｋｉｎ、ＴＤＺ或ＣＰＰＵ都不能诱导芽的分化,而ＭＳ附加玉米素能有效地诱导芽分化,其中以２．０ｍｇ／Ｌ玉米素效果最好。     

巴戟天组织培养和快速繁殖研究

以巴戟天顶芽及嫩茎节段为外植体,以MS为基本培养基,通过不同的激素种类和浓度配比,建立巴戟天组培快繁体系。结果表明,外植体表面消毒以70%酒精预处理60s,再用0.1%HgCl2浸泡10min,效果较好,茎节为外植体优于顶芽。培养基MS+BA1.0mg/L+IBA0.05mg/L利于诱导出芽,可用于初代培养;MS+BA1.0mg/L+IBA0.2mg/L利于形成丛生芽,用于继代增殖,繁殖系数6.0/50d;1/2MS+IBA0.4~0.8mg/L适宜诱导生根获得再生植株,生根率100%;生根苗移栽于排水良好的火土或砂土中,成活率90%。     

Induction of Continuous Tepal Differentiation from in Vitro Regenerated Flower Buds of Hyacinthus orientalis

Continuous differentiation of tepals was successively induced from regenerated flower buds in Hyacinthus orientalis L. cv. White Pearl by controlling the exogenous hormones and explant ages. In 250 days of subculture, each flower bud differentiated an average of more than 70 tepals, with a maximum of over 140 tepals. Studies on the morphogenesis and characteristics of growth and development of the flower buds indicate that the first whorled organ of the flower bud was perianth which consisted of perianth tube and tepals grown at the top of the perianth tube, which is the same as the flower bud of the wild type in H. orentalis. The second and third whorls of the flower bud, which should be stamen and pistil in the wild type, but remained as the tepals in the regenerated flower bud. Growth of the regenerated flower bud was faster in the first several months of culture, then slowed down gradually with time. After 150 days in culture the flower bud growth and organ differentiation became very slow. Other than the tepal differentiation the regenerated flower buds also differentiated at random positions some small flower buds that also differentiated the tepals only. Histological observation revealed that the origin of the regenerated flower buds was jointly participated by some cells in the epidermal and subepidermal layers at the inner surface of the perianth explant, and the inner small flower buds were originated from the meristem which was formed by the transformation of the parenchyma at the base of the very young tepal. The authors also compared and discussed the similarities and differences of the phenotypes between the regenerated flower bud in Hyacinthus and agamous flower in Arabidopsis, from which, they have hypothesized on the role of the hormones in the promotion and termination of the gene expressions by an order of development in plant.