多效唑对水稻未成熟胚愈伤组织诱导,分化和壮苗培养的影响

多效唑(Mult-Effect Triazole,MET)是一种良好的植物生长延缓剂,具有多方面的生理效应。已在水稻、小麦、油菜等作物中应用。本文报道了多效唑对水稻未成熟胚愈伤组织诱导、分化以及对壮苗培养的影响,为植物试管苗的正常化生产提供了有效的化学调控措施。1.材料早粳“T53”未成熟胚,愈伤组织,再生绿芽;早籼87-5-0,“湘竹443”再生绿芽。多效唑由本所生理系提供。     

高羊茅组织培养再生体系及GUS基因瞬间表达研究

以成熟种子为外值体,对高羊茅纰织培养和植株再生体系进行了优化,分析了不同浓度2．4-D、6-BA和激动素对高羊茅愈伤组织诱导和愈伤组织分化成苗的影响．结果表明：9．0mg／L 2．4-L)对愈伤组织的诱导效果最佳．0．2mg／L激动素是愈伤组织分化成苗的最适浓度．二者的诱导率和分化率分别达到68．08%和45．83％。在愈伤组织继代培养基中附加1．0mg／L 2．4-D、0．5mg／L 6-BA和1．25mg／L CuSO4;有利于胚性愈伤组织的形成,可以明显促进愈伤组织分化。同时．采用基因枪法将GUS基因导入高羊茅愈伤组织中,通过组织化学染色检测到了GUS瞬间表达活性;并对影响CUS基因瞬间表达的因素进行了分析．以期为提高基因枪法遗传转化效率提供参考。     

多效唑（MET）对啤酒花试管苗生长和移栽的影响

在培养基中加入0.1-0.2mg/L浓度的多效唑（MET）,啤酒花试管苗植株矮壮,单株生根数量增加、移栽成活率提高13％。0．5－2．0mg/L时,试管苗基部愈伤组织增多,枝叶失绿,生长受到抑制。初步发现多将唑对啤酒花试管苗的效力与葡萄糖用量呈正相关。     

Cytological and Embryological Studies on Haploid Plant Production from Cultured Unpollinated Ovaries of Nicotiana tabacum L.

We obtained mature haploid (n = 24) ovary plants from in vitro cultured unpollinated young ovaries. These ovaries were induced to form embryoids which then developed into plants. The results obtained are summarized as follows: 1. The origin of development of the ovary haploid plants has been followed by light microscopy. Embryological abservations revealed that there are two ways of plantlet production: (1) Ovary haploid plant was derived from the macrospore without an intervening callus phase. (2) Ovary haploid plant was derived directly from the egg cell of mature embryo sac. In addition, Callus derived haploid plant was also obtained from the base and the tip of a bud of the above mentioned haploid plantlet. In same medium embryoids was derived from callus. Finally, plantlet was developed. 2. The exogenous hormones are necessary for high induction frequency of embryoid from unpollinated isolated young ovary, but these are not definitely necessary for induction of embryonic callus to form embryoids which then developed into plant. 3. The induction frequency of embryoid from in vitro cultured ovary and embryonic callus significantly increased when the concentration of thiamine, pyridoxine, ascorbic acid, nicotinic acid, inositol and folic acid was raised.     

Embryogenic Callus Formation and Plantlet Regeneration of Piragmites communis

The present paper reports the embryogenic callus formation and plantlet regeneration of Phragmites communis. The results have been obtained as follows: The efficiency of callus induction was much higher, if reed seeds were used as explants. No dedifferentiation was observed by using leaf sheath and leaf blade as explants. The optim, um concentration of sucrose was 4% in medium. VB group and inositol had beneficial effects on callus growth. But yeast extract inhibited callus induction and callus growth markedly. For this inhibited reaction, the higher concentration, the more obviously the callus growth was inhibited. Higher levels of 2,4-D had unfavourable effects on callus growth in callus subculture. The concentration of 2,4-D in dedifferentiation medium had relation to embryogenic callus formation. Embryogenic callus had higher frequency of differentiation for long-term subculture. On the other hand, nonembryogenic callus most often lost their morphogenetic competence. Authors found that the surface structure of the two types of calluses was different by means of observation by scanning electron microscope. The peroxidase and the esterase isoenzyme pat- terns, as well as the soluble protein of both types of calluses were different too.     

源于小孢子培养的大麦耐盐变异体获取

以大麦品种‘花30’作为供试材料,比较了甲基磺酸乙酯（EMS）和平阳霉素处理小孢子60Co γ-射线辐照处理离体穗和干种子,对300mg·L-1NaCl胁迫培养下游离小孢子的愈伤组织产量和愈伤组织在0.3％NaCl胁迫筛选下的绿苗产量的影响。结果表明,EMS处理离体小孢子和60Co γ-射线辐照千种子的愈伤组织产量和绿苗产量明显优于平阳霉素处理小孢子和60Co γ-射线辐照离体穗。以16份源于种子辐照处理的再生植株自交一代种子为供试材料,比较了在0．3％NaCl胁迫下种子的发芽率和幼苗的成活率以及植株的分蘖数、株高和单株产量。结果表明,‘花30’发芽率为0,供试的16份耐盐变异体中,有14份材料在NaCl胁迫下的发芽率优于‘花30’,鉴定出4份耐盐性明显优于‘花30’的变异体材料。选择耐盐变异体作为供试材料,测定了变异体中Na＋／H＋逆向转运蛋白基因NHXl、NHX2和NHX3和编码甜菜碱醛脱氢酶（BADH）的两个同工酶基因曰肋,和BBD2的表达模式和表达量,结果表明变异体耐盐性的提高与这些基因的表达量存在联系。     

芦苇胚性愈伤组织的形成及植株的再生

以芦苇种子为外植体,其愈伤组织的诱导率最高。叶鞘和叶片不发生脱分化。培养基中最合适的蔗糖浓度为4%。维生素 B 类、肌醇对愈伤组织的生长起促进作用。而酵母提取物对愈伤组织的诱导和生长具有明显的抑制作用。这种抑制效应,将随酵母提取物浓度的提高而增大。愈伤组织的继代培养,随培养基中2,4-D 浓度的提高,其平均鲜重明显降低。脱分化培养基中2,4-D 浓度对胚性愈伤组织的诱导形成具有一定的相关性。胚性愈伤组织经30代继代培养依然具有90%的分化频率,只是每块愈伤组织的分化苗数减少。反之,非胚性愈伤组织则完全丧失形态发生的能力。对两类愈伤组织进行扫描电镜的观察,发现其表面结构有很大差异。其过氧化物酶、酯酶同工酶谱以及可溶性蛋白的含量均有明显的差别。     

Characterization of factors affecting plantlet regeneration from rice (Oryza sativa L.) callus

Various components of culture media were tested to characterize factors affecting plantlet regeneration from rice (Oryza sativa L.) callus. It was found that plantlet regeneration from rice callus was affected by concentrations of gelling agents, osmoticum, and the combination of hormones in the regeneration medium. High concentrations (4–6 g/l gellan gum, 10–16 g/l agar) of gelling agents promoted regeneration frequency. However, the total number of plantlets decreased with gellan gum concentrations above 4 g/l. Addition of sorbitol (15–75 g/l) promoted plantlet regeneration. However, the addition of mannitol was inhibitory and no regeneration was observed at concentrations above 30 g/l. This difference in the effects on regeneration suggests that sorbitol had another function besides as a osmoticum. High regeneration frequency was obtained with combinations of NAA (0.05–0.5 g/l) and kinetin (0.5–2 mg/l). However, higher concentrations (2 mg/l) of NAA are preferred to increase the total number of regenerated plantlets.     

曼陀罗茎段愈伤组织诱导和再生植株的研究

本试验以曼陀罗茎段为外植体,在附加不同植物激素组合的培养基中对愈伤组织的诱导和植株再生进行研究。结果表明:采用修改的MS培养基(除去甘氨酸,维生素B1含量增加至0.5mg/L,pH5.5)附加2mg/L2,4-D可由曼陀罗茎段诱导大量胚性愈伤组织;愈伤组织继代选用0.5mg/L2,4-D为宜;不定芽的诱导采用MS培养基(20g蔗糖,8g琼脂,0.1g水解干酪素) 6-BA(0.5mg/L);幼苗进一步转接至1/2MS IBA(0.2mg/L)生根培养基中,可完成曼陀罗茎段愈伤组织诱导和再生植株的组织培养过程。     

水稻抗氨基酸类似物突变体的筛选——Ⅰ.抗S-(2-氨乙基)L-半胱氨酸(AEC)突变体的筛选

以赖氨酸类似物S-(2-氨乙基)L-半胱氨酸(AEC)为选择剂,从水稻花药培养中筛选出一个抗性突变体(R_(AEC))。突变体愈伤组织经过6个月继代培养后仍保持抗性稳定。R_(AEC)再生植株根尖诱导的愈伤组织经过3个月继代培养也保持稳定的抗性。R_(AEC)细胞内赖氨酸含量提高了近2倍,苏氨酸提高5倍多。其他氨基酸,如蛋氨酸、酪氨酸、丝氨酸等都有较大量的提高。 R_(AEC)愈伤组织对赖氨酸加苏氨酸混合物也具有抗性。突变体植株较原始类型稍矮小,巳正常结实。     

芒不同基因型愈伤组织诱导及分化的差异

芒(Miscanthus sinensis)具较高的生物学产量, 是一种极具发展前景的纤维素类能源植物。以芒的8种不同基因型幼穗为外植体, 进行了组织培养研究。结果表明, 不同基因型芒在愈伤组织诱导率、胚性愈伤组织诱导率和胚性愈伤组织分化率等方面均存在显著差异。W89和W70均具较高的愈伤组织诱导率, 分别为91.7%和89.1%; W69的外植体几乎全部褐化, 且未能诱导出愈伤组织。W89、W70和W17的胚性愈伤组织分化率较高, 达50%以上。另外, 发现愈伤组织诱导率与细胞壁木质素含量间呈显著的负相关。该研究建立了稳定且有效的再生体系, 并初步确定W89和W70可作为芒组织培养的理想材料, 为芒的遗传转化、定向改良和良种快繁提供了技术支持。     

NAA、IBA和PP333对怀山药试管苗生长发育的影响

就不同生长调节剂对怀山药试管苗生长发育的影响进行了系统的研究。结果表明 :( 1 )NAA、IBA和PP3 3 3 均能诱导根的生成 ,但根形成的时间、发生方式及发达程度均不相同。低浓度的NAA( 0 .1～ 0 .5mg.L 1)有利于根的生成 ,但高浓度的NAA(≥ 2mg .L 1)则易形成愈伤组织 ,且随着浓度的升高 ,愈伤组织化程度变大 ,根多来源于愈伤组织 ;IBA( 0 .1～ 2mg .L 1)对根的生成较为有利 ,其中以IBA 1mg .L 1的生根效果较好 ;PP3 3 3 ( 0 1～ 8mg.L 1)有利于根的生成 ,根形成的最早、最多 ,且随着浓度的升高 ,根更加粗壮发达。 ( 2 )PP3 3 3 抑制试管苗的纵向伸长生长 ,使株高降低 ,但却显著地促进了根系的发育 ,使试管苗生长健壮 ,叶色浓绿 ,叶片增多。这种效应随着PP3 3 3 浓度的升高而加强。从培养壮苗的角度来看 ,PP3 3 3 ( 2～ 4mg.L 1)是最佳浓度     

Stem Apex Culture and Quality Improvement of Tubercles of Pinellia ternata

Formation of Plantlets was achieved when stem apex of Pinellia ternata Brier. Cultured in vitro on MS medium with KT 0. 5 mg/L + NAA 0.2 mg/L (MSI). With petioles of the plantlet as explants callus could be induced after cultured for a week on MS medium with 2, 4-D 2.0 mg/L + KT 0.5 mg/L (MSII). Calli were subcultured once in every month. After 3--4 months a kind of friable calli could be selected, from which the tubercles could be differentiate and the plantlets formed when transfered onto MSI. But before callus differentiation, a lot of roots were formed on callus. The plantlets could be produced directly from the petiole segment. It was found that the stem growing tip was always covered by the leaf primordium and the former leaf primordium was covered by the latter leaf primordium during the differentiation of the apical bud of tubercle. The frenquency of plantlet differentiation from callus and petioles was over 70%. The rate of regeneration of plantlet on liquid static culture was twice as much as that on solid culture. All plantlets grew well after being transfered into the plot. The fresh weight of tuber-plant was 103 % higher than that of control (cultivated plant come from tubers). The alkaloid content of tubers come from tuberplant was 0. 344%, that of control was 0. 203% and 0. 264% for the wild tuber.     

Correlation of epiphyllous bud differentiation with foliar senescence in crassulacean succulent Kalanchoe pinnata as revealed by thidiazuron and ethrel application

Leaves of Kalanchoe pinnata have crenate margins with each notch bearing a dormant bud competent to develop into a healthy plantlet. Leaf detachment is a common signal for inducing two contrastingly different leaf-based processes, i.e. epiphyllous bud development into plantlet and foliar senescence. To investigate differentiation of bud and its correlation, if any, with foliar senescence, thidiazuron (TDZ), having cytokinin activity and ethrel (ETH), an ethylene releasing compound, were employed. The experimental system was comprised of marginal leaf discs, each harbouring an epiphyllous bud. Most of the growth characteristics of plantlet developing from the epiphyllous bud were significantly inhibited by TDZ but promoted by ETH. The two regulators modulated senescence in a manner different for leaf discs and plantlet leaves. Thus, TDZ caused a complete retention whereas ETH a complete loss of chlorophyll in the leaf discs. In contrast, the former resulted in a complete depletion of chlorophyll from the plantlet leaves producing an albino effect, while the latter reduced it by 50% only. In combined dispensation of the two regulators, the effect of TDZ was expressed in majority of responses studied. The results presented in this investigation clearly show that the foliar processes of epiphyllous bud differentiation and senescence are interlinked as TDZ that delayed senescence inhibited epiphyllous bud differentiation and ETH that hastened senescence promoted it. A working hypothesis to interpret responsiveness of the disc-bud composite on lines of a source-sink duo, has been proposed.     

In Vitro Culture of Different Explants from Hubei Photoperiod Sensitive Genic Male-Sterile Rice

Young panicles, immature embryos, stem nodes, stem tips, leaf segments, root tips and anthers from Hubei Photoperiod Sensitive Genie Male-Sterile Rice (Oryza sativa subsp, japonica) “Nongken 58” (abbr. 58s) and fertile “Nongken 58” (abbr. 58f) were examined for callus induction, plant regeneration and direct plantlet formation on differentiation medium. 58s and 58f had equal ability in all explants cultures except anther cultures. The induction frequency of the anther callus and the regeneration frequency of the green plant in 58s were much lower than those in 58f, and such differences were not affected markedly by the change of fertility of 58s donor plants. Young panicle, immature embryo, stem node, stem tips showed direct plantlet formation when cultured on differentiation medium containing NAA and kinetin. Different explants produced various types of responses. Young panicles could produce callus and then regenerate plantlets. Evidences from histological observation showed that the plant regeneration in direct plaatlet formation of young panicles were mainly organogenetic, bowever, somatic embryogenesis was also possible.     

多效唑连用其它植物激素对水稻试管苗生长的影响

本实验采用继代多年的花培体细胞无性系Hu18再生绿芽(0.5mm)为起始材料,研究多效唑(MET)与其它激素配合使用对试管苗的调控作用,结果指出:(1)单独使用MET对绿芽生长有毒害作用,除2,4-D、GA,外,MET与适宜浓度的其它激素配合使用才能发挥增苗、壮苗作用,其中以MET与BA配合使用的培养效果最好,MET与NAA,C_2H_4配合使用的效果次之;(2)MET与其他激素配合使用不但能降低植株高度,促进根系发育,而且可以延长试管苗的保存时间;(3)MET与乙烯利配合使用能加速绿芽成苗速度,而与其他激素配合使则延缓绿芽成苗速度,如与2,4-D配合使用则延缓2,4-D对绿芽的脱分化进程;(4)在本实验条件下,以MET 2.5mg/L+BA 2mg/L+NAA 0.2mg/L配合使用有利根芽的协调生长。本文还从植株干物质累积,叶细胞结构,细胞活力等方面进行了探讨。     

盾叶薯蓣花药培养及单倍体植株的获得

以处于单核期的盾叶薯蓣花药为材料,经愈伤组织途径成功获得单倍体盾叶薯蓣新材料。对盾叶薯蓣花药培养、植株再生过程的研究表明:来自不同居群的外植体对花药愈伤组织诱导有显著影响;W14是适合花药愈伤组织诱导的基本培养基;花药愈伤组织增殖和分化的适宜培养基为:MS基本培养基 BA2.0mg/L IAA0.2mg/L;生根培养基为添加了IBA2.0mg/L、NAA0.4mg/L和0.5g/L活性碳的MS培养基。以流式细胞仪和根尖染色体压片法检查花培植株的结果表明,有8%～12%的个体为单倍体。实验结果为进一步开展单倍体育种及相关理论研究提供了材料和技术基础。     

The role of salicylic acid and carrot embryogenic callus extracts in somatic embryogenesis of naked oat (Avena nuda)

Enhanced somatic embryogenesis and plant regeneration have been obtained using young leaf bases of naked oat (Avena nuda) as explants by including salicylic acid (SA) and carrot embryogenic callus extracts (CECE) in media. A 5- and 4-fold improvement was achieved in somatic embryogenesis and plant regeneration on the corresponding media supplemented with 0.5 mM SA and CECE as compared to control, respectively. Some physiological and biochemical changes were assayed in both embryogenic callus (EC) and non-embryogenic callus (NEC). The results indicated that superoxide dismutase activity was stimulated and catalases and ascorbate peroxidase activities were inhibited, while the O₂ ^- (superoxide anion) content was reduced and the hydrogen peroxide level was promoted in EC compared with NEC. Reduced malondialdehyde content and relative electrolyte leakage were also detected in EC.     

Induction Of Callus and Regeneration of Plantlets in Stem Segment Culture of Chinese Gooseberry

The stem segments of male and female plants of Actinidia chihensis Planch. var. chinensis C. F. Liang and A. chihensis Planch. var. hispida C. F. Liang were cultured in vitro. Callus and plantlets were obtained on MS basic medium supplemented with BAP (0.5, 2 ppm) + IAA (0.05 ppm) or zeatin (0.1–10 ppm). Best results were obtained on medium added with 1 ppm zeatin. It was possible to induce callus in both species but the frequency of callus redifferentiating into plantlet in A. chinensis Planch. var. hispida was far higher than that in A. chinensis Planch. var. chinensis and the frequency of plantlets derived from the female plant was higher than those from the male plant. The stem of plantlet was cut off when it reached 1 cm in height, then soaked in the solution with IBA 50 ppm for 2 hr. or with IBA 2 ppm for 24 hr. and the excised stem was transferred onto medium with 1/2 MS+ 1% sucrose +0.5% active carbon for rooting. It is possible that this technique of obtaining whole plants by stem segment culture provides a method for thes multiplication of female plants and the good individual plants of chinese gooseberry.     

Genetic Study of the Rice Embryo Culture In Vitro

The embryo culture in vitro response was examined among ten rice (Oryza sativa L.) cultivars and 26 cross combinations to evaluate the correlation between callus induction rate and differentiation rate with plantlet regeneration rate, the influence of parents to hybrid Fl embryo culture in vitro as well as the cytoplasmic effects. Plantlet regeneration rate was used as the product of callus production and regeneration capacity. The ten pure-lines, the five F1s and their reciprocal hybrid as well as the ten F1s among the ten lines were evaluated for callus production and regeneration capaticy. Significant variation was observed among the 36 genotypes in callus induction rate, callus differentiation rate and plantlet regeneration rate on embryo culture in vitro. The positive correlation between general callus induction rate and differentiation rate with plantlet regeneration rate was significant. There was a similar trend for callus induction rate between maternal parents and Fis during mature embryo culture in vitro. However, parent-offspring correlation for callus differentiation rate and plantlet regeneration rate were nonsignificant. Whether cytoplasmic effects for embryo culture response exist among the six pure-lines was examined py the differences between reciprocal F1 hybrids. The extent of cytoplasmic effects depended on cross combination.