In vitro propagation through axillary bud multiplication in different mulberry genotypes

Axillary buds from 5 genotypes of mulberry belonging to 4 species were cultured on modified MS basal medium. A total of 30 media combinations were tried for all the genotypes. The response of axillary buds and the requirement for growth regulators varied with genotype. In Morus indica BAP (0.25–0.5 mg/l), and in M. alba and M. rotondifolia GA₃ (0.5–1.0 mg/l)were found to induce sprouting. Two genotypes of M. bombycis, namely Schimanochi and Mizusawa, developed healthy shoots on the incorporation of 2,4-D (0.5–1.0 mg/l) and BAP (0.5–2.0 mg/l), respectively. IBA (0.5 mg/l), along with cytokinin/auxin/gibberellin, had no effect on bud growth but helped root induction. Shoots developed from the axillary buds were further multiplied as nodal explants. MS basal medium supplemented with 0.5 mg/l IBA and LS vitamins was found best to produce healthy plantlets in all the genotypes. An average 89% survival was observed on transferring the plantlets to soil.Abbreviations MS Murashige and Skoog (1962) - LS Linsmaier and Skoog (1965) - IBA 3-indole-butyric acid - GA₃ Gibberellic acid - BAP 6-Benzylaminopurine - Kn Kinetin - 2,4-D 2,4-Dichlorophenoxyacetic acid     

Micropropagation of a fruit tree, Morus australis Poir. syn. M. acidosa Griff.

High frequency bud break and multiple shoots were induced in nodal explants collected between November to February from a 5 year old tree of Morus australis Poir syn. M. acidosa Griff. on Murashige and Skoog's medium supplemented with 6-benzylaminopurine (1.0 mg/1). Incorporation of gibberellic acid (0.3 mg/l) along with BAP (1.0 mg/l) not only induced faster bud break from nodal explants as well as from apical shoot buds, but it also enhanced the frequency of bud break. Nodal explants were more responsive than apical shoot buds. The shoots formed in vitro were multiplied further as nodal segments, and an average multiplication rate of 6-fold per subculture was established within 4–5 months. The shoots were successfully rooted on half-strength MS containing a combination of indole-3-acetic acid, indole-3-butyric acid and indole-3-propionic acid, each at 1.0 mg/1. The plantlets were successfully hardened off and established in natural soil.Abbreviations BAP 6-benzylaminopurine - GA₃ gibberellic acid - KN kinetin - IAA indole-3-acetic acid - IBA indole-3-butyric acid - IPA indole-3-propionic acid - MS Murashige and Skoog (1962) medium - NAA 1-naphthalene acetic acid     

Effects of external ABA,GA<Subscript>3</Subscript> and NAA on the tiller bud outgrowth of rice is related to changes in endogenous hormones

Hormones play an important role in regulating the growth of rice tiller buds. However, little is known about the hormonal changes that occur during tiller bud growth and the mechanism of hormonal regulation of tiller bud growth. Here, two rice cultivars, Yangdao 6 (Indica) and Nanjing 44 (Japonica), were used to investigate the changes in plant hormones during tiller bud growth and the mechanism that underlies the hormonal regulation of tiller bud growth. In the present study, panicles were removed after heading to stimulate the growth of dormant tiller buds located at the elongated upper internodes. At the same time, external abscisic acid (ABA), gibberellic acid (GA₃) and α-naphthalene acetic acid (NAA) were applied. The results demonstrated that auxin and cytokinin (CTK) play important and different roles in the regulation of tiller bud growth. Auxin in the nodes inhibits tiller bud growth, while CTK is transferred to the tiller buds to promote growth. The inhibitory effects of GA₃ and NAA on tiller bud growth are mainly due to the control of the indole-3-acetic acid (IAA) or CTK contents in plants. As opposed to auxin and CTK, the ABA contents in nodes and tiller buds remained unchanged before tiller bud growth after panicle removal. Meanwhile, external ABA application only slightly slowed the growth of the tiller buds, suggesting that ABA may not be a key regulator of tiller bud growth. These results indicate that auxin, CTK and ABA together likely play roles in the regulation of tiller bud growth.     

Effect of Gibberellic Acid and Monophenols on Flowering of lmpatiens balsamina in Relation to the Number of Inductive and Non-inductive Photoperiodic Cycles

A single treatment of plants with GA₃ (gibberellic acid) is not adequate to cause induction under LD (long day: 24-h photo-period) condition, but its effect is added to the sub-threshold induction caused by one SD (short day: 8-h photoperiod) cycle. Floral bud initiation is hastened, and the number of floral buds and flowers per flowering plant increases in plants receiving a single treatment with the combination GA₃+ SA (salicylic acid) accompanying a single SD cycle. However, the increase on 10 replicate basis is more marked in plants receiving three treatments with the combination GA₃+β-N (β-naphthol) and five treatments with the combination GA₃+ SA accompanying six and 10 SD cycles, respectively. The number of floral buds and flowers decreases with an increase hi the number of SD cycles, but it is higher in plants treated with GA₃, SA or GA₃+β-N than in the water-treated controls. — Under long days, treatment of plants with the combinations GA₃+ SA or GA₃+β-N accelerates the initiation as well as increases the number of floral buds. While a minimum of five treatments with GA₃ or of 25 with SA or β-N alone is needed for floral bud initiation under a 24-h photoperiod, three treatments are adequate to induce floral buds with the combination GA₃+ SA or GA₃+β-N under continuous illumination. Ten or more treatments with these combinations under a 24-h photoperiod produce more flowers than the same treatments under an 8-h photoperiod.     

Plant regeneration through somatic embryogenesis from mature leaf explants of Eryngium foetidum,a condiment

Eryngium foetidum L. is an important plant cultivated as a leafy vegetable and for its essential oil, which are of high economic value in international trade market. Plants were regenerated through somatic embryogenesis from mature leaf explants of field grown plants. Leaf explants produced dark brown, compact callus on Linsmaier and Skoog (LS) medium with the combination of 1.0 mg l^-1 2,4-dichlorophenoxy acetic acid (2,4-D) and 1.0 mg l^-1 benzylaminopurine (BAP). Somatic embryos were induced from embryo-forming callus cultures on Murashige and Skoog (MS) medium supplemented with 0.1 mg l^-1 2,4-D, 2.0 mg l^-1 BAP and 1.0 mg l^-1 gibberellic acid (GA₃). Subsequently, conversion of these somatic embryos into plantlets occurred on MS medium supplemented with 1.0 mg l^-1 GA₃ and/or 0.1 mg l^-1 BAP. The regenerated shoots were rooted and elongated on MS medium supplemented with 0.1 mg l^-1 IAA and 1.0 mg l^-1 GA₃. These plantlets were hardened and transferred to the soil. This revised version was published online in June 2006 with corrections to the Cover Date.     

枣花芽分化过程中营养物质和内源激素含量及抗氧化酶活性变化研究

以新疆主栽品种灰枣和骏枣的花芽为材料,测定不同分化时期花芽的可溶性糖、还原糖、淀粉、可溶性蛋白含量,SOD、POD、PPO、CAT活性以及内源GA3、IAA、ABA、ZT水平的变化,并分析它们与花芽分化的关系,为枣花芽分化调控提供理论参考.结果表明:(1)灰枣和骏枣花芽可溶性糖、还原糖和淀粉含量在花芽分化过程的变化趋势...     

Rapid clonal propagation of three mulberries,Morus cathayana HemsL,M. lhou Koiz. andM. serrata Roxb., through in vitro culture of apical shoot buds and nodal explants from mature trees

High-frequency bud break and multiple shoots were induced in apical shoot buds and nodal explants ofMorus cathayana, M. lhou andM. serrata on Murashige and Skoog (MS) medium containing 0.5–1.0 mg/l 6-benzylaminopurine (BAP). Addition of gibberellic acid (0.4 mg/l) along with BAP induced faster bud break both in apical shoot buds and nodal explants and also enhanced the frequency of bud break in all three species. Shoot culture initiation was greatly influenced by explant type, explant age and explanting season. The shoots were successfully rooted on half-strength MS medium containing a combination of indole-3-acetic acid, indole-3-butyric acid and indole-3-propionic acid, each at 1.0 mg/l. The plantlets were successfully acclimated and eventually established in soil.Abbreviations BAP 6-Benzylaminopurine - GA ₃ Gibberellic acid - IAA Indole-3-acetic acid - IBA Indole-3-butyric acid - IPA Indole-3-propionic acid - Kn Kinetin - MS Murashige and Skoog (1962) medium - NAA 1-Naphthalene acetic acid     

Regulation of flower development in cultured ears of maize (Zea mays L.)

Summary Young ears of maize were cultured in two different liquid media containing either kinetin (KN) or kinetin + gibberellic acid (KN + GA₃) in order to manipulate stamen and gynoecium development. In KN medium, stamens developed and gynoecia aborted in the flowers of the cultured immature ears. In the KN + GA₃ medium, however, ovaries with silks developed and stamens aborted. These differential morphological events were recorded with SEM photomicrographs at regular intervals after excision of ear inflorescences. In addition, the mitotic activity in the developing or aborting organs was determined over a 75-h period. It increased from 6% to 14% in developing organs (i.e. stamens in KN medium, and gynoecia in KN + GA₃ medium) and gradually decreased to 1% in the degenerating organs (i.e. gynoecia in KN medium, and stamens in KN + GA₃ medium) by 45 h of culture. The mitotic activity reached zero in degenerating flower organs by 75 h of culture. Whether these differential sensitivities to the exogenously applied members of these two plant growth regulator classes are unique to our in vitro system or reflect a more general control feature of in vivo inflorescences must await further clarification.     

Dwarfism Caused by Floral-Bud Removal in Petunia and its Reversal by Gibberellin

The effect of floral-bud removal at different stages of developmenton the plant height and on the total number of buds of Petuniawas studied. Continuous removal of all the floral buds 2 d beforeanthesis caused a marked decrease in plant height and also increasedthe total number of floral buds formed thereafter. At otherstages of floral bud development, bud removal had a lesser effecton both phenomena. Moreover, the plants did not respond to budremoval at anthesis. GA₃ at 25 ppm applied to plants from which the buds had beenremoved, promoted stem elongation. The most pronounced effectwas on plants from which the buds were removed 2 d before anthesis,but it had no effect on plants from which the buds were removedat anthesis stage. The possible involvement of endogenous growth hormones in theresponse of Petunia plants to floral-bud removal and to applicationof GA₃ is discussed. Bud removal, bud number, dwarfness, GA₃, Petunia, plant height     

Effect of Gibberellic Acid and Phosfon on the Critical Dark Period Reqnirement for Flowering of Impatiens balsamina cv. Rose

The critical dark period requirement for flowering of Impatiens balsamina L. cv. Rose, an obligate short day plant, is about 8.5 hours. While GA₃ completely substituted for the dark period requirement, Phosfon prolonged it to 9.5 hours. GA₃ hastened and Phosfon delayed the initiation of floral buds under all photoperiods. Floral buds opened into flowers only during 8 and 14 hour photoperiods in control and Phosfon-treated plants but during all photoperiods in GA₃-treated ones. The delay in floral bud initiation and flowering was correlated with shifting up of the node bearing the first floral bud and flower respectively. While GA₃ increased the numher of floral buds and flowers in all photoperiods except 8-hour, Phosfon increased their number in the 14-hour photoperiod only. The number of flowering plants decreased with increasing photoperiod regardless of GA₃ and Phosfon application. The effect of Phosfon was completely or partially overcome, depending upon the photoperiod, by simultaneous application of GA₃.     

Influence of benzyladenine and gibberellic acid on organogenesis in lsquo;Crimson Giantrsquo; Easter cactus

Experiments were performed to determine the influence of gibberellic acid (GA₃) and benzyladenine (BA) on organogenesis of lsquo;Crimson Giantrsquo; Easter cactus [Hatiora gaertneri (Regel) Barthlott] phylloclades cultured in vitro. The numbers of flower buds and new phylloclades increased linearly as BA concentration increased from 0 to 444.1 micro;M. GA₃ increased the number of new phylloclades when present in moderate concentrations (2.9 or 28.9 micro;M), but inhibited flower bud formation when present in concentrations as low as 0.3 micro;M. The inhibitory effect of GA₃ on flower bud formation was diminished when the medium was amended with BA at 44.4 or 444.1 micro;M. Explants cultured in media that contained 288.7 micro;M GA₃ produced fewer organs (new phylloclades plus flower buds) compared to those cultured in media with 0, 0.3, 2.9, or 28.9 micro;M GA₃. BA and GA₃ concentrations also affected the percentage of explants with flower buds and the percentage of explants with new phylloclades. This study shows that organogenesis in H. gaertneri can be controlled by varying the concentrations of BA and GA₃ in the culture medium.     

Factors influencing direct shoot regeneration from mature leaves of Jatropha curcas, an important biofuel plant

In order to establish a highly efficient and sustainable regeneration system, we systematically researched the key factors affecting direct shoot regeneration from Jatropha curcas leaves that were collected from Hainan (HN1-1), Lijiang (LJ3-1), and Yuxi (YX2-12) provinces in China. The L₉(3⁴) orthogonal test of thidiazuron (TDZ), kinetin (Kn), and gibberellic acid (GA₃) were studied, and the explant type, growth age, and cultivar of leaves were subsequently investigated. Simultaneously, the combinations of plant growth regulators (PGRs) promoting shoot bud proliferation, elongation, and root establishment were examined. The results showed that the best medium for shoot bud induction was Murashige and Skoog (MS) medium supplemented with 1.0 mg/L TDZ, 0.5 mg/L Kn, and 0.5 mg/L GA₃. TDZ was the key PGR, while Kn and GA₃ played an important role in shoot bud elongation and the number of shoots per leaf disk, respectively. The induced shoot buds proliferated and readily elongated in MS medium with 0.3 mg/L 6-benzylaminopurine and 0.01 mg/L indole-3-butyric acid (IBA) and established roots in half-strength MS medium supplemented with 2.0 mg/L IBA. Using the previously described methods, the third to fifth leaves were found to be the best explant source for shoot bud induction, with a high induction rate, large shoot numbers per disk, excellent proliferation, and consistent rooting. With the use of this regeneration system, the shoot bud induction rate increased from the reported rate of 53.5% to more than 90% using different explants and cultivars, and the shoot number per leaf disk (shoot length?≥?0.5 cm) increased from 1.6 to 3.5. Thus, this optimized regeneration system will effectively promote the propagation and genetic transformation of J. curcas.     

Effects of Abscisic Acid on Phenolic Content and Lignin Biosynthesis in Tobacco Tissue Culture

A significant depression of callus growth resulted from low concentrations of abscisic acid (ABA) added to the medium recommended by Linsmaier and Skoog. Low concentrations also decreased the chlorogenic acid and lignin content of the callus, and generally decreased amounts of scopolin and scopoletin in the tissue. Gibberellic acid (GA₃) stimulated callus growth in a low concentration (0.1 mg/1) and inhibited growth at a high concentration (10.0 mg/1). Both levels of GA₃ increased scopoletin accumulation in tobacco callus. A high concentration of GA₃ increased the accumulation of scopolin and chlorogenic acids, whereas a low concentration decreased the amounts of these two phenolic compounds. In comparison with the control, lignin synthesis was stimulated by a low GA₃ concentration, but a high GA₃ concentration did not have a significant effect. Both low and high concentrations of GA₃ overcame ABA inhibition of growth and lignin synthesis, and partially reversed ABA inhibition of scopoletin production. However, GA₃ did not reverse the inhibitory effect of ABA on scopolin production. The low concentration of GA₃ overcame the inhibition of chlorogenic acid production resulting from a 0.01 mg/1 concentration of ABA, but this was the only reversal of chlorogenic acid inhibition resulting from addition of GA₃ to the medium.     

Micropropagation of wild beet (Beta maritima) from inflorescence pieces

In order to investigate the regeneration of wild beet (Beta maritima) from inflorescence pieces, the effects of growth regulator, genotype, explant source and stage of plant development on adventitious shoot formation and rooting in vitro and subsequent transplanting in the glasshouse were tested. Inflorescence tips produced more adventitious shoots than sub-apical segments and the best micropropagation was achieved on a Murashige and Skoog (MS) medium supplemented with 1.0 mg l^–1 BAP. Addition of auxin was not beneficial. The induction rate of adventitious shoots was genotype-dependent and influenced by the stage of plant development. Adventitious shoots were produced from the base of the flower buds, i.e. from the receptacle, not from axils or stalks and only a few buds on inflorescence tip explants produced adventitious shoots. Rooting was increased by using a MS medium with 3% sucrose supplemented with 1.0 mg l^–1 NAA. There was no variation in leaf morphology of the transplants. This work shows that inflorescence tips can be used successfully as explants for in vitro multiplication of sugar beet and wild beet.Abbreviations BAP benzylaminopurine - IBA indole-3-butyric acid - GA₃ gibberellic acid - MS Murashige and Skoog medium - NAA naphthaleneacetic acid Author for correspondence     

In vitro propagation of shoot buds of Carica papaya L. (caricaceae) var. Honey Dew

Shoot buds from the saplings and the fruit bearing plants of Carica papaya L.. var. Honey Dew (papaya) initially treated with Gentamycin were cultured in modified MS media, each with a different hormonal combination, for the establishment of cultures and multiplication and rooting of plants. About 43% of explants from fruit bearing plants and 69% of those from saplings remained free of contamination and retained regeneration capacity when treated in 500 mg/l Gentamycin. For the establishment of the explants a medium containing 1 mg/l GA₃ and 2 mg/l kinetin was necessary. When established buds were transferred to medium containing 1 mg/l NAA and 3 mg/l kinetin, calli were initiated at cut ends of shoot buds; multiplication started on transfer to NAA (0.1 mg/l) and BAP (0.5 mg/l) medium. Cultures have been maintained for the last twenty months without any loss in multiplication rate. Rooting was induced in medium with reduced salt concentration containing 2 mg/l IBA. Shoot elongation was induced after prolonged culture in the same rooting medium.Abbreviations MS Murashige and Skoog, 1962 - SH Schenk and Hildebrandt, 1972 - GA₃ Gibberellic acid - Kn Kinetin - NAA Napthaleneacetic acid - BAP 6 -Benzylaminopurine - IBA Indole-3-butyric acid - IAA Indole-3-acetic acid     

In vitro propagation of the medicinal herbs Ocimum americanum L. syn. O. canum Sims. (hoary basil) and Ocimum sanctum L. (holy basil)

A procedure is outlined for in vitro propagation of two medicinal herbs, Ocimum americanum L. syn. O. canum Sims (hoary basil) and Ocimum sanctum L. (holy basil), using axillary shoot buds. Multiple shoot formation was induced from shoot bud explants of both species on Murashige and Skoog medium (MS) supplemented with benzyladenine (BA). The optimum BA concentrations for shoot proliferation were 0.25 mg/l for O. americanum and 1.0 mg/l for O. sanctum. Incorporation of 0.5 mg/l gibberellic acid (GA₃) along with BA in the culture medium resulted in a marked increase in the frequency of axillary branching as well as multiple shoot formation. Shoot buds collected between September through December were most responsive in culture. Shoots of O. americanum were rooted on half-strength MS supplemented with 1.0 mg/l indole-3-butyric acid (IBA), whereas O. sanctum rooted best on medium with 1.0 mg/l naphthaleneacetic acid (NAA). The plantlets were hardened off and successfully established in natural soil, where they grew and matured normally.Abbreviations BA N⁶-benzyladenine - 2,4-D 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - IAA indole-3-acetic acid - IBA indole-3-butyric acid - MS Murashige and Skoog (1962) medium - NAA 1-naphthaleneacetic acid     

High frequency embryoid and plantlet formation from tissue cultures of the Finger millet-Eleusine coracana (L.) Gaertn

Compact nodulated embryogenic callus differentiated from cultured seeds of Eleusine coracana (Finger Millet) on Murashige and Skoog (1962) basal medium with 2,4-dichlorophenoxyacetic acid (1.0, 3.0 mg ^l). This embryogenic callus was maintained on a medium with a lower level of 2,4 — dichlorophenoxyacetic acid. At every subculture the embryogenic callus had some preexisting embryoids in it. With this method of subculture the callus has retained its morphogenic potential for four years. Following transfer to media with different levels of auxins and cytokinins, the callus showed varied patterns of growth and morphogenesis. Embryoids could be germinated in profusion to form plantlets which could be transferred to the field. Shoot buds also differentiated from the whole surface of the embryoid or from the flattened meristemoids.Abbreviations BAP 6-benzylaminopurine - IAA indole-3-acetic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - NAA napthaleneacetic acid - IBA indolebutyric acid - KN kinetin - MS Murashige and Skoog (1962) - GA₃ Gibberellic acid     

FLOWER CULTURE OF A MALE STERILE STAMENLESS-2 MUTANT OF TOMATO (LYCOPERSICON ESCULENTUM)

Young floral buds of a male sterile stamenless-2 (sl-2/sl-2) mutant of tomato were cultured, at the sepal primordia stage, in a liquid Murashige and Skoog medium containing either benzylaminopurine (BAP) or gibberellic acid (GA₃) or both. In the basal medium (BM), the buds initiated petal and stamen primordia only and they showed limited development. In buds grown in BM supplemented with 10^–6 M BAP, all types of organ primordia were initiated but the petals remained small and the stamens and carpels were immature. Well-developed flowers with a normal complement of floral organs were, however, produced in a medium containing both BAP (10^–6 M) and GA₃ (10^–7 M to 10^–5 M). The development of stamens was variable and ranged from the complete absence of microsporogenesis to the formation of abnormal pollen. Gynoecium development was normal and ovules with megaspores were produced in the ovary. The results show that male sterility in the sl-2/sl-2 mutant can be expressed in vitro and that GA₃ is essential for the in vitro growth and development of all the floral organs of this mutant.     

In vitro propagation of Populus x wilsocarpa — a hybrid of ornamental value

Populus x wilsocarpa, a hybrid of important ornamental value, cannot be seed-propagated, nor grafted, since a compatible rootstock has not been identified. A micropropagation protocol consisting of a series of steps was therefore developed to facilitate the commercial production of this species. The technique involved the transfer of swelling buds to a growth initiation medium with the following composition: N6 macronutrients, MS micronutrients and vitamins supplemented with 0.5 mg l^-1 BAP. The best buds were from dormant twigs, stored at 0–2°C and then forced to burst prior to culture initiation. Shoot multiplication was on a basal WPM medium including 0.1 mg l^-1 BAP and 0.001 mg l^-1 NAA. Shoot elongation and rooting was also on a basal WPM medium supplemented with 1.0 mg l^-1 GA₃ followed by a transfer to a peat-perlite mix in the greenhouse.Abbreviations ABA abscisic acid - BAP benzylaminopurine - GA₃ gibberellic acid (GA₃) - MS Murashige and Skoog [17] - NAA naphthaleneacetic acid - N6 medium [Chu et al., 7] - WPM woody plant medium [16]     

Effect of exogenous gibberellic acid,abscisic acid,and benzylaminopurine on epicotyl dormancy of cultured herbaceous peony embryos

Epicotyl dormancy was broken in cultured peony (Paeonia lactiflora Pall.) embryos after topical application of agarose gels containing gibberellic acid, with optimum growth at 1.5 mM gibberellic acid. Addition of 100 M abscisic acid to the medium resulted in complete inhibition of gibberellic acid-stimulated promotion of dormant epicotyls. Epicotyl dormancy was also broken in embryos by culture on media containing 1 or 10 M benzylaminopurine. A highly significant increase in leaf number occurred when embryos were both cultured on medium containing benzylaminopurine and treated topically with gibberellic acid. Anatomical and morphological studies indicated that the increase in shoot growth was due to the development and growth of 1) buds formed at the cotyledonary node, 2) axillary buds, and 3) adventitious meristems originating from subepidermal parenchymatous tissue.Abbreviations ABA abscisic acid - BA N⁶-benzylaminopurine - DMSO dimethyl sulfoxide - GA₃ gibberellic acid - LS Linsmaier and Skoog