外源激素对脱毒马铃薯扦插苗生长及生理效应研究

以‘早大白’马铃薯脱毒苗为试验材料,通过人工温室苗床栽培探讨外源激素萘乙酸(NAA)、吲哚丁酸(IBA)和硼酸生根处理对扦插脱毒苗生长和生理特征的影响,为马铃薯微型薯的实际生产提供理论依据。结果显示:(1)外源激素生根处理较对照扦插苗根系长势好,根活力、根系可溶性蛋白含量增加。(2)外源激素生根处理较对照脱毒扦插苗光合速率提高、叶绿素含量增加,光合物质的形成与积累增多。(3)外源激素生根处理较对照叶片SOD、POD、CAT活性和游离氨基酸含量均表现出不同程度的上升,MDA含量下降,衰老减缓。(4)外源激素生根处理有利于小区产量和单株结薯数的增加。研究表明,不同外源激素生根处理可改善脱毒马铃薯扦插苗农艺性状和生理指标,并以NAA 100mg/L+IBA 50mg/L+硼酸17.5mg/L配方处理植株的长势最好,叶绿素含量、保护酶活性及游离氨基酸含量最高,净光合速率大、小薯膨大速度快且单株结薯数量及产量增加显著,更利于发挥脱毒薯的增产优势。     

多肉植物离体再生研究进展

通过品种、外植体类型、培养基、外源植物激素的选择等几方面对国内多肉植物的离体再生研究现状进行了综述。发现多肉植物的离体再生研究多以种子、叶片、茎段、花器官等为外植体;不定芽诱导阶段常用的培养基类型为MS培养基,不定根诱导常用1/2 MS培养基;6-苄氨基腺嘌呤(6-BA)、萘乙酸(NAA)和激动素(KT)作为常见的外源植物激素,被广泛运用于多肉植物愈伤组织诱导、不定芽诱导、不定根诱导等离体再生各阶段;分析了当前存在的问题和发展前景,以期为日后多肉植物的相关研究提供可行性建议。     

Tissue cultures of Artemisia pallens: Organogenesis,terpenoid production

Germinated seedlings of Artemisia pallens gave three types of cultures on MS medium supplemented with different plant growth hormones. Medium containing BA+2,4-D stimulated unorganized callus; BA+IAA medium, semi-organized tissues interspersed with shoot buds; and BA+NAA+IAA medium, multiple shoot cultures. The in vitro shoots developed roots in medium devoid of growth hormones. TLC and GLC analysis of the tissue extracts showed that linalool was present in the cultured tissues, with maximum concentration in the unorganized tissue. Although the TLC profiles of the three culture extracts were similar, the extracts did not contain the major polar compounds of the plant. The plant extracts contained more polar compounds and gave the characteristic fragrance of davana.Abbreviations MS Murashige & Skoog's basal medium - BA benzyladenine - Kn kinetin - NAA naphthaleneacetic acid - IAA indoleacetic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - PCV packed cell volume     

Characterization of a novel putative zinc finger gene MIF1: involvement in multiple hormonal regulation of Arabidopsis development

Phytohormones play crucial roles in regulating many aspects of plant development. Although much has been learned about the effects of individual hormones, cross-talk between and integration of different hormonal signals are still not well understood. We present a study of MINI ZINC FINGER 1 (MIF1), a putative zinc finger protein from Arabidopsis, and suggest that it may be involved in integrating signals from multiple hormones. MIF1 homologs are highly conserved among seed plants, each characterized by a very short sequence containing a central putative zinc finger domain. Constitutive overexpression of MIF1 caused dramatic developmental defects, including dwarfism, reduced apical dominance, extreme longevity, dark-green leaves, altered flower morphology, poor fertility, reduced hypocotyl length, spoon-like cotyledons, reduced root growth, and ectopic root hairs on hypocotyls and cotyledons. In addition, 35S::MIF1 seedlings underwent constitutive photomorphogenesis in the dark, with root growth similar to that in the light. Furthermore, 35S::MIF1 seedlings were demonstrated to be non-responsive to gibberellin (GA) for cell elongation, hypersensitive to the GA synthesis inhibitor paclobutrazol (PAC) and abscisic acid (ABA), and hyposensitive to auxin, brassinosteroid and cytokinin, but normally responsive to ethylene. The de-etiolation defect could not be rescued by the hormones tested. Consistent with these observations, genome-scale expression profiling revealed that 35S::MIF1 seedlings exhibited decreased expression of genes involved in GA, auxin and brassinosteroid signaling as well as cell elongation/expansion, and increased expression of ABA-responsive genes. We propose that MIF1, or the protein(s) with which MIF1 interacts, is involved in mediating the control of plant development by multiple hormones.     

Growth Hormones and Propagation of Cymbidium in vitro

Protocorms of Cymbidium (Orchidaceae) were grown on solid or liquid medium with macro-nutrients according to Wimber (van Raalte 1967) and iron, micro-nutrients and vitamins according to Nitsch (1968) the medium also contained 2% sucrose. The effects of 1) the auxins; indol-3yl-acetic acid (IAA), α-naphthaleneacetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D); 2) the cytokinins; 6-furfurylaminopurine (kinetin) and benzyladenine (BA) and 3) the gibberellin; gibberellic acid (GA) were examined alone or in combinations. IAA had no effect alone. NAA resulted in optimal fresh weight at 10 μM and the protocorms were vigorous, but lighter green than usual. 2,4-D caused a high weight increase at 1 μM, but the protocorms were abnormal. Higher concentrations of NAA and 2,4-D inhibited chlorophyll synthesis. On solid medium kinetin (100 μM) induced a growth of many small shoots, but had no effect on the fresh weight. In liquid medium, kinetin promoted a callus formation and fresh weight increase. BA had effects similar to kinetin, but at lower concentrations. GA alone promoted shoot and leaf growth. Combinations of kinetin and NAA resulted in a maximal fresh weight increase at kinetin concentrations one tenth of the NAA concentrations. The optimal growth and the best development occurred at 10 μM NAA and 1 μM kinetin. NAA and kinetin together could limit the shoot and leaf growth induced by GA.     

One step shoot tip multiplication and rooting of Digitalis thapsi L.

Shoot tips from seedlings of Digitalis thapsi L. were cultured on Murashige and Skoog's medium and the effect of various auxins (2,4-D, NAA and IAA) were analyzed alone or in combination with cytokinis (BA and kinetin). Shoot multiplication and direct rooting of the new shoots were obtained after four weeks of culture in MS medium without hormones, but callus formation and the appearance of abnormal phenotypes were frequent. The addition of auxins to the cultures prevented the formation of callus but not the appearance of variant phenotypes. Both drawbacks could be avoided by combination of NAA or IAA with BA or kinetin. The best results for shoot multiplication and direct rooting were obtained with 0.5 mg l^-1 NAA and 0.1 or 0.5 mg l^-1 kinetin.Abbreviations BA 6-benciladenine - 2,4-D 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid - Kin kinetin - NAA naphtalene acetic acid - MS Murashige and Skoog     

In vitro propagation of Podophyllum peltatum L. by the cultures of embrya and divided embrya

Excised embrya and subsequently divided embrya of Podophyllum peltatum were cultured on the Murashige and Skoog medium supplemented with different growth regulators, because traditional methods of breaking seed dormancy failed. The growth of excised embrya was stimulated by 1 or 0.1 mg dm-3gibberellic acid (GA3), 0.1 mg dm-3 GA3 + 0.2 mg dm-3 kinetin (kin), or 0.2mg dm-3 kin. GA3 (1 mg dm-3) showed the best effect; after 5 weeks the plantlets had 1.5 - 2 cm long cotyledons, 5 - 6 cm long roots, 88 % of embrya germinated and developed further. The addition of 0.5 mg dm-3 zeatin + 0.2 mg dm-3 naphtaleneacetic acid (NAA), 0.2 mg dm-3 NAA, and 1 mg dm-3 kinetin inhibited the growth of embrya. 1 mg dm-3 kinetin + 0.1 mg dm-3 NAA, 0.1 mg dm-3 zeatin and 0.2 mg dm-3 6-benzylaminopurine resulted in a compact appearance of plantlets and a lower germination rate. Divided embryo cultures produced plantlets via somatic embryogenesis which occurred only on the 2,4-dichlorophenoxyacetic acid containing media. The maturation of somatic embrya was observed on media without any auxin. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of Chilling on DNA Endoreplication in Root Cortex Cells and Root Hairs of Soybean Seedlings

Relative nuclear DNA contents in cortex parenchyma cells in root segments of 3- and 7-d-old soybean seedlings grown at 25 °C and in plants grown for 3 d at 25 °C, and then for 4 d at 10 °C, were determined with cytophotometry. Measurements revealed that in each variant the cortex cell nuclei with DNA content between 2C and 8C were in all the examined segments and nuclei with 8C – 16C DNA appeared in higher parts of roots. However, in chilled plant cells the number of 8C – 16C DNA nuclei was very low. Therefore, chilling inhibited endoreplication in comparison with plants grown at 25 °C for 7 d, and even reduced endopolyploidy level as compared to the initial seedlings, i.e. 3-d-old plants. DNA contents in root hairs grown at 25 °C (control) and in root hairs emerged at 10 °C were also determined. In controls 4C – 8C DNA nuclei predominated while in chilled plants an additional population of 2C – 4C DNA appeared. Thus a reduction of DNA synthesis was brought about by low temperature. The occurrence of an intermediate DNA contents besides those with full endoreplication cycles suggests the possibility of differential DNA replication. This suggestion seems to be supported by the lack of ³H-thymidine incorporation into root hair nuclei at the examined developmental stage both in control and chilled root hairs. The same number, but larger, chromocentric lumps in polyploid cortex cell nuclei of higher root zones, in comparison to meristematic nuclei, suggests that endoreduplication process occurred. This revised version was published online in July 2006 with corrections to the Cover Date.     

THE INFLUENCE OF GROWTH SUBSTANCES ON THE INDUCTION OF APOGAMY IN PTERIDIUM GAMETOPHYTES

Experiments employing sequences of three media demonstrated the effect of growth substances on the induction of apogamy. The most effective sequence was 4% sucrose 1–14 days, 4% sucrose plus growth substance 15–28 days, and 0.1% sucrose 29–56 days. In this sequence concentrations of NAA, IAA, and GA promoted apogamous shoot formation. A higher NAA concentration than optimal for shoot formation stimulated apogamous root formation in all medium sequences. Kinetin was without effect or inhibitory to apogamy. Combination of kinetin/GA or GA/NAA concentrations did not increase the apogamous response. One combination of the kinetin/NAA concentrations had a synergistic effect on the apogamous shoot formation. Additions of GA to the synergistic kinetin/NAA combination had an antagonistic effect on the apogamous response.     

HORMONE-INDUCED ENDOREDUPLICATION PRIOR TO MITOSIS IN CULTURED PEA ROOT CORTEX CELLS

One-mm-thick cortical explants excised aseptically from 10-11 mm behind the tip of 3-day-old roots of the garden pea, Pisum sativum, cv. ‘Little Marvel’ were cultured on a synthetic nutrient medium supplemented with auxin or auxin and cytokinin. Nuclear DNA contents were measured in cells of the explants at the outset and at specified times during culture up to seven days. Fixed and sectioned preparations were stained with the Feulgen method using the DNA-specific dye auramin-O. Fluorescent microspectro-photometric measurements of individual nuclei were made from each cortical population. At day zero all cortical nuclei measured were either 2c or 4c with respect to their DNA content. In the presence of the auxins, indoleacetic acid and 2,4-dichlorophenoxyacetic acid, and the cytokinin, kinetin, DNA values increased to multiples of the 2c level with populations at the 8c and 16c level predominating after three days of culture as well as at seven days. In the presence of auxins alone no change in DNA values was observed during three days. Kinetin concentrations as low as 0.01 ppm were already effective. The data are interpreted to show that cytokinin, in the presence of auxin, induces two rounds of DNA synthesis prior to the first mitoses, the first round being connected with chromosome doubling by endoreduplication and the second one with normal mitosis. From this we inferred that tetraploid cells in leguminous root nodules might have arisen in the same way, i.e., by endoreduplication prior to the first mitoses induced by the rhizobial division stimulus, unless the chromosome number of root cortical cells had already been doubled by endoreduplication in the normally differentiating root systems.     

Improved transient production of a cellulase enzyme in detached sunflower leaves using plant hormones

A synthetic endoglucanase E1 was transiently expressed in detached whole sunflower leaves using recombinant Agrobacterium tumefaciens. As a means to increase protein accumulation, the effect of plant hormones on E1 production was investigated using six different hormones: methyl jasmonic acid (JA), gibberellin A3 (GA), indole acetic acid (IAA), 2,4-dichlorophenoxyacetic acid (2,4-D), kinetin (CK), and salicylic acid (SA). Among the six hormones, JA was best at boosting E1 concentrations, increasing yields as much as six-fold and the JA treatment was especially effective at an early phase of leaf incubation likely by enhancing T-DNA transfer at an early leaf incubation phase. Finally, co-infiltrating combinations of two different hormones revealed an antagonistic interaction between JA and SA in A. tumefaciens-mediated transient expression.     

Growth and stomata development of Arabidopsis hypocotyls are controlled by gibberellins and modulated by ethylene and auxins

The plant hormones gibberellin (GA), ethylene and auxin can promote hypocotyl elongation of Arabidopsis seedlings grown in the light on a low nutrient medium (LNM). In this study, we used hypocotyl elongation as a system to investigate interactions between GA and ethylene or auxin and analysed their influence on the development of stomata in the hypocotyl. When applied together, GA and ethylene or auxin exerted a synergistic effect on hypocotyl elongation. Stimulated cell elongation is the main cause of hypocotyl elongation. Furthermore, hypocotyls treated with GA plus either ethylene or auxin show an increased endoreduplication. In addition, a small but significant increase in cell number was observed in the cortical cell files of hypocotyls treated with ethylene and GA together. However, studies with transgenic seedlings expressing CycB1::uidA genes revealed that cell division in the hypocotyl occurs only in the epidermis and mainly to form stomata, a process strictly regulated by hormones. Stomata formation in the hypocotyl is induced by the treatment with either GA or ethylene. The effect of GA could be strongly enhanced by the simultaneous addition of ethylene or auxin to the growth medium. Gibberellin is the main signal inducing stomata formation in the hypocotyl. In addition, this signal regulates hypocotyl elongation and is modulated by ethylene and auxin. The implication of these three hormones in relation to cell division and stomata formation is discussed.     

枸杞的组织培养及植株再生的条件优化

目的:探讨枸杞组织培养及其植株再生条件的优化。方法:应用MS培养基为基本培养基,以各种不同激素配比进行枸杞愈伤诱导、分化诱导及根的诱导。结果:以枸杞叶片为外植体,利用2,4-D(2,4-二氯苯氧基乙酸)与KT(细胞分裂素)不同配比诱导出了愈伤组织。利用6-BA(6-苄基腺嘌呤)与NAA(α-萘乙酸)不同浓度的配比组合,成功地进行了杞再生芽诱导及根系诱导。结论:以MS培养基为基本培养基,并采用各种激素的不同配比,可以优化枸杞植株的再生条件。     

<Emphasis Type="Italic">In vitro</Emphasis> organogenesis and antioxidant enzymes activity in <Emphasis Type="Italic">Acanthophyllum sordidum</Emphasis>

The effect of various hormonal combinations on callus formation and regeneration of shoot and root from leaf derived callus of Acanthophyllum sordidum Bunge ex Boiss. has been studied. Proteins and activity of antioxidant enzymes were also evaluated during shoot and root organogenesis from callus. Calli were induced from leaf explants excised from 30-d-old seedlings grown on Murashige and Skoog medium containing 4.52 μM 2,4-dichlorophenoxyacetic acid + 4.65 μM kinetin. Maximum growth of calli and the most efficient regeneration of shoots and roots occurred with 2.69 μM 1-naphthalene acetic acid (NAA), 2.69 μM NAA + 4.54 μM thidiazuron and 2.46 μM indole-3-butyric acid. Protein content decreased in calli and increased significantly during regeneration of shoots from callus. Superoxide dismutase activity decreased in calli comparing to that of seedlings, then increased in regenerated shoots and roots. High catalase activity was detected in seedlings and regenerated shoots, whereas high peroxidase activity was observed in calli and regenerated roots.     

Isolation and properties of arginase from a shade plant, ginseng (Panax ginseng C.A. Meyer) roots.

Arginase (EC 3.5.3.1) was purified to homogeneity from root tissues of three-year-old ginseng (Panax ginseng C.A. Meyer), shade plant, and was found to be an extraordinarily large molecule relatively stable to heat. The enzyme was decameric having a molecular mass of 352,000 Da, with an optimal temperature and pH of 60 degrees C and 9.5, respectively. Analogues of arginine could not replace it as substrate, and a cysteine residue is at or near the active site. Maximum activity was obtained with Mn(2+) and Co(2+) also activated the proteins, whereas, both agmatine and 5'-deoxy-methylthioadenosine were inhibitors. Specific activities of the enzyme in sliced ginseng roots were increased by plant hormones such as GA(3), IAA, kinetin and putrescine, whereas the activities of the purified enzyme were unaffected by putrescine. Increases in arginase activities by these plant hormones could affect metabolism of polyamine intracellularly.     

The Effect of Growth Hormones on Cell Division and Expansion in Liquid Suspension Cultures of Acer pseudoplatanus

The effects on cell division and cell size of indole-3-aceticacid (IAA), gibberellic acid (GA), and kinetin were studiedin liquid suspension cultures of cambial cells derived fromAcer pseudoplatanus. It was shown that all three hormones promotecell division and that the effects of both GA and kinetin areadditive to those of IAA, but the effects of GA and kinetintogether are not additive. Treatment with IAA resulted in anincrease of mean cell size (indicating that cell expansion ispromoted), but after GA or kinetin treatment the mean cell sizewas smaller, indicating that little cell expansion had takenplace after each division. The results are discussed in relationto previous work on the effects of hormones in the intact cambiumand to current theories on the interactions of growth hormones.     

An excellent source of vegetative buds for use in plant hormone studies on apical dominance

When studying the role of plant hormones in the control of growth at apical meristems, it is often difficult to obtain needed amounts of physiologically uniform buds. A source and method are described for obtaining sufficient quantities of large, uniform buds and for the treatment of the buds with indoleacetic acid and kinetin. Buds from the root system of Euphorbia esula L. were grown in Petri plates, with agar suspending the short root sections from which they emanate. Plant hormones are applied by their incorporation in the agar. The effect of various concentrations of indoleacetic acid and kinetin on bud growth was examined.     

Recovery of loss in chlorophyll and 2,6-dichlorophenol indophenol Hill reaction of isolated chloroplasts during dark induced aging of intact maize seedlings

Loss in the content of pigments and decline in the efficiency of thylakoid membranes to reduce 2,6-dichlorophenol indophenol (DCPIP) have been investigated during dark induced senescence of attached leaves of maize seedlings. The chlorophyll degradation during senescence is differentially inhibited by indoleacetic acid (IAA), gibberellic acid (GA) and kinetin. IAA and GA behave as mild senescence inhibitors in comparison to kinetin. However, in comparison to light, kinetin is relatively less efficient in counteracting senescence. Dark-induced loss in chlorophyll content is fully recovered by light when the dark incubation period is relatively short. The pattern of light recovery of loss in photoelectron transport during dark-aging is similar to the recovery kinetics of chlorophyll. Continuous kinetin treatment of dark-incubated seedlings inhibits the chlorophyll degradation but with decreased duration of kinetin treatment, the efficiency of the hormone to inhibit chlorophyll loss is reduced. The kinetin-induced inhibition of pigment loss is small in comparison with the effect of light.