Rapid propagation of Cucurbita pepo L. by culture of meristem tips

A tissue culture technique has been developed for the rapid multiplication of pumpkin (Cucurbita pepo L.) clones. Meristem-tips from seedlings of cultivar ‘Cinderella’ were grown initially on MS medium containing 2.56 mg l^?1 Kinetin and 8 mg l^?1 IAA, and then transferred to experimental media. Maximum shoot proliferation occurred on MS medium containing 1 mg l^?1 BA and no auxin. Cultures were rooted after 2–3 weeks on MS medium containing 8 mg l^?1 IAA and no cytokinin.     

Effect of gibberellic acid (GA3) on elongation and rooting of ‘St. Julien A’ rootstock in vitro

‘St. Julien A’ (Prunus instititia L.) rootstock was induced to proliferate shoots on a modified half-strength Murashige and Skoog (MS) medium. Cultures treated with 12.5 mg l^?1 gibberellic acid (GA₃) produced elongated shoots suitable for rooting. Elongated shoots were placed in media with indolebutyric acid (IBA) or indole-3-acetic acid (IAA) with or without a 16-day dark incubation. Light (16-h photoperiod) inhibited rooting. IAA (4 mg l^?1) was ineffective in promoting rooting. Rooting was best when shoots were incubated in the dark with IBA (4 mg l^?1). GA₃ was deleterious to shoots, causing chlorosis and apical die-back. Light regime interacted with auxin treatments in affecting shoot condition. Shoot condition was better on shoots treated with IBA and dark-incubated; while those treated with IAA were better when light-incubated.     

In vitro propagation of GF677 hybrid rootstock (Prunus persica × Prunus amygdalus) from mature cotyledons

Adventitious shoot regeneration from mature cotyledons of GF677 rootstock (Prunus persica × Prunus amygdalus) was achieved in vitro. Thidiazuron (N-phenyl-N-1,2,3-thidiazol-5-yl-urea; TDZ) at 32 µM gave the highest percentage of cotyledons forming adventitious shoots (68.8%) and the highest number of shoots per cotyledon (4.8) on Quoirin and Lepoivre (QL) medium. On QL medium containing 32 µM TDZ, exposure of the proximal segments of cotyledons to darkness at the start of culture increased the percentage of cotyledons forming adventitious shoots (62.5%) when compared with those kept under light conditions (15%). A combination of 0.72 µM gibberellic acid and dark treatment resulted in at least 2.7-fold more elongated shoots than non-treated shoots. The highest rooting percentage (100%) occurred on 0.5× Murashige and Skoog medium supplemented with Gamborg (B5) vitamins and 2 mg l^?1 indole-3-butyric acid. Rooted plantlets were acclimatised under greenhouse conditions with a 70% survival rate.     

Effects of growth regulators on growth and flowering in Hippeastrum hybridum hort

Soaking of bulbs in 3 concentrations of indoleacetic acid (IAA), gibberellic acid (GA₃), 2-chloroethyltrimethyl ammonium chloride (cycocel) or 2-chloroethylphosphonic acid (ethrel) showed various responses on growth and flowering. IAA increased the weight and number of bulblets, GA₃ increased bulb weight. Cycocel (1000 mg l^?1) increased the number of flowers, while GA₃ increased the diameter of the flowers.Application of IAA at 100 mg l^?1 and GA₃ at 10, 100 or 1000 mg l^?1 twice as foliar spray at an interval of 30 days promoted the number of bulblets on the treated plants, while high concentrations of cycocel and ethrel (1000 mg l^?1) increased the weight of bulblets. All concentrations of IAA, GA₃ and 1000 mg l^?1 cycocel increased the number and size of the flowers.     

Rapid propagation of white cabbage by tissue culture

A tissue culture technique has been devised to produce plants of white cabbage (Brassica oleracea v. capitata L.f. alba) from heads stored at 0.5 ± 0.5°C for 8 months. Meristem-tips (0.5–2 mm diameter), excised from heads of 11 accessions, were grown initially on MS medium containing 2.56 mg l^?1 of kinetin and then induced to proliferate shoots on MS medium with 12.8 mg l^?1 kinetin. Subsequent transfer to a kinetin-free medium resulted in root development in 1–2 weeks. Rooted plantlets were readily established in soil. Plantlets obtained in this way from parent cabbages containing turnip mosaic virus remained infected.     

Influence of growth regulators on setting,retention and weight of fruits in two cultivars of litchi

Investigations were undertaken to explore the possibility of improving setting, retention and weight of fruits in ‘Early Seedless’ and ‘Calcuttia’ cultivars of lichi (Litchi chinensis) by means of growth regulators. Indole acetic acid (IAA) at 20, 40 and 80 mg l^?1, 2,4-dichlorophenoxy acetic acid (2,4-D) at 2,4 and 8 mg l^?1 and gibberellic acid (GA₃) at 50, 100 and 150 mg l^?1 were sprayed on panicles in the first fortnight of April, when 50–100% flowers had opened. All 3 growth regulators caused a favourable effect on fruit setting, fruit retention and weight of individual fruits, but IAA at 20 mg l^?1 proved the best for enhancing setting, GA₃ at 50 mg l^?1 for increasing retention and GA₃ at 100 mg l^?1 for improving fruit weight. IAA and GA₃ should, therefore, be used in combination. Between the 2 cultivars tested, ‘Calcuttia’ proved superior to ‘Early Seedless’ in fruit setting, fruit retention and weight of individual fruits.     

Callus and axillary-bud culture of Vitis vinifera ‘Sylvaner Riesling’

Healthy growth of serially subcultured callus of the grape Vitis vinifera cultivar ‘Sylvaner’ was obtained by incubation at 30° C in continuous light in a defined culture medium containing 2% w/v sucrose, 1.0 mg l^?1 1-naphthaleneacetic acid (NAA) and 0.2 mg l^?1 kinetin (K). Organogenesis was not induced in this callus by alteration in the absolute or relative levels of NAA and K.Continued shoot initiation was obtained by culture of axillary buds in a medium containing 10^?5 M Benzyladenine (BA). Plantlets could be generated from these shoot buds by transfer to media containing 10^?7 M BA or lacking a cytokinin.     

In vitro regeneration from mature leaf explants of Pelargonium rapaceum (L.) L’Hérit

Methods to regenerate whole plants from mature leaf explants of Pelargonium rapaceum (L.) L’Hérit were established. To optimize shoot induction, leaf explants were cultured on media containing different types and combinations of plant growth regulators. Growth was initiated within 17–24 days culture, and included callus formation, and root or shoot organogenesis ranging from 20 to 100% regeneration. Shoots were induced only when explants were cultivated on MS medium containing a combination of NAA and kinetin, NAA and BAP, IAA and Kinetin, or IAA and BAP. On media containing NAA and BAP, dark incubation was critical for efficient direct shoot regeneration from explants. Direct shoot formation and the highest number of shoots per explant (17.6) were obtained from leaf explants cultured in the dark for 30 days on MS medium containing 0.1 mg l⁻¹ NAA and 0.1 mg l⁻¹ BAP. Shoots cultured on MS medium containing 0.1 mg l⁻¹ NAA formed tuberous roots with microtubers within 42 days. Healthy regenerated plants were acclimated and transferred to a greenhouse.     

Influence of high concentrations of cytokinins on the production of somatic embryos by germinating seeds of tomato,aubergine and pepper

Summary

Somatic embryos of tomato, aubergine and pepper were initiated from intact seedlings when seeds were cultured on medium containing 6-benzylaminopurine (BAP) or thidiazuron (TDZ). The percentage of explants producing somatic embryos was highest for aubergine on media containing low concentrations of BAP, i.e. 0–10 mg l^–1, for tomato at 15–20 mg l^–1 and for pepper at 40–80 mg l^–1. Aubergine and tomato produced fewer somatic embryos per responsive seedling when cultured on medium containing TDZ, and pepper did not produce any somatic embryos on media containing 0–20 mg l^–1 1 TDZ. Morphogenesis of the seedlings producing somatic embryos was similar for all the genotypes, i.e. the seedlings were dwarf, only the cotyledons expanded, development of the apical meristem and the root were suppressed and a ring-like crown of nodular tissue developed at the base of the hypocotyl from which somatic embryos were initiated. Co-cultivation of tomato and aubergine seeds with seeds of pepper in media containing 0, 5, 10, 15 and 20 mg l^–1 BAP inhibited somatic embryogenesis in tomato and aubergine instead of assisting somatic embryogenesis in pepper. This is discussed in relation to the recent findings for the induction of somatic embyrogenesis in peanut (Arachis hypogea L.) and the role of BAP and TDZ.     

In vitro regeneration of Camellia reticulata by somatic embryogenesis

Camellia reticulata L. plantlets were regenerated by direct and indirect somatic embryogenesis from immature zygotic embryos. Initial explants (cotyledon sections and embryonic axes) produced somatic embryos without intermediate callus tissue when grown on Murashige and Skoog’s basal medium with 30 gl^-1 sucrose and no growth regulators; the somatic embryos completed their development in 4-6 weeks in the same medium. Embryogénie competence was increased by 0.5 and 1 mg l^-1 IBA. Histological observation showed the embryos to originate from epidermal and subepidermal cells of the cotyledon and hypocotyl explants. Secondary somatic embryos developed directly from the cotyledons and hypocotyl region of primary somatic embryos by a process that was morphologically very similar to that occurring on zygotic explants. Direct repetitive embryogenesis was maintained by this system. Up to 40% germination occurred when mature somatic embryos were isolated and incubated in medium supplemented with 1 mgl^-1 GA₃ + 1 mgl^-1 IAA. Indirect somatic embryogenesis was induced in callus differentiated on cotyledon explants after three months’ culture in media containing IBA or NAA and/or BAP, embryogenic capacity being retained by callus subcultured on 0.5 mg l^-1 IBA + 1 mg l^-1 BAP.     

Physiological response of cauliflower in relation to manganese-boron interaction

Summary

Cauliflower (Brassica oleracea L. var. botrytis) was grown in refined sand at low (0.0011 mg l^?1), normal (0.55 mg l^?1) and excess (5.5 mg l^?1) Mn, each at three levels of B, deficient (0.0033 mg l^?1), normal (0.33 mg H) and excess (3.3 mg l^?1). In Mn deficient cauliflowers a deficiency of B accentuated visible symptoms of Mn deficiency and aggravated the Mn deficiency effects i.e. a decrease in dry matter, leaf Mn, sugars, starch, chlorophyll, Hill reaction and specific activity of aldolase and an increase in the concentrations of proline and inorganic phosphorus. In cauliflowers exhibiting B toxicity symptoms under conditions of excess B, excess Mn increased leaf Mn, sugars, nucleic acids, protein P, nucleic acid P, acid-labile P and leaf B. Excess Mn decreased the concentrations of DNA, protein nitrogen, chlorophyll and activities of peroxidase, aldolase and leaf B, leaf Mn in boron deficient cauliflowers.     

In vitro propagation of ‘Troyer’ citrange from epicotyl segments

Isolated epicotyl, root meristem and root segment tissues of ‘Troyer’ citrange [Poncirus trifoliata (L.) Rat. × Citrus sinensis (L.) Osbeck] were established in continuous culture to compare their regeneration potential. Callus was obtained from these explants on a Murashige—Skoog (MS) medium containing NAA (10 mg l^?1) and BAP (0.1–10 mg l^?1). Formation of shoots from root segments was direct without callus formation on MS medium containing BAP (10 mg l^?1) and NAA (1 mg l^?1). Shoot formation from epicotyl callus occurred on MS medium containing 0.25 mg l^?1 BAP and 0.1 mg l^?1 NAA. Formation of shoots from epicotyl segments occurred on MS medium containing BAP (0.5 mg l^?1) and NAA (0.1–1.0 mg l^?1), while rooting of regenerated shoots occurred in treatments containing 2.0 mg l^?1 NAA alone. This system provides a rapid method for propagation of ‘Troyer’ citrange.     

The influence of wax emulsion,morphactin and gibberellic acid on the storage behaviour of indian gooseberry fruits

Qualities of Indian gooseberry fruits (Phyllanthus emblica L.) were determined after dip-treatment with wax emulsion (WE) with or without morphactin (Mor) and gibberellic acid (GA₃). Dip-treatments with 100–500 mg l^?1 Mor reduced marketability by inducing browning and high weight loss, but loss in ascorbic acid was checked and phenols increased. 10 mg l^?1 Mor maintained marketability at par with control, effectively controlled loss in ascorbic acid, and increased acidity and reducing sugars. GA₃, although failing to control the loss of ascorbic acid, was effective in checking browning and thereby increased the market value of fruits as compared with controls and Mor-treated fruits.WE with or without Mor (10 mg l^?1) controlled browning, accumulation of phenols and losses in weight and moisture as compared with 10 mg l^?1 Mor, 100 mg l^?1 GA₃, or control, but could not retain ascorbic acid in comparison to 10 mg l^?1 Mor-treated fruits. Marketability of fruits having had treatment with either WE or GA₃ was the same. Fruits having had combined treatment with WE and 10 mg l^?1 Mor had maximum marketability. Minimum marketability was observed in fruits subjected to a combined treatment of WE + 100 mg l^?1 GA₃ due to maximum infection with Aspergillus spp. and Penicillium spp.     

无子西瓜郑抗4号的体细胞胚诱导及植株再生(英文)

以无子西瓜(CitrulluslanatusMansfeld)郑抗4号无菌苗子叶为外植体,进行了体细胞胚发生及植株再生的研究,结果表明,无菌苗应先进行3d暗培养,然后采取光照16h/d和黑暗8h/d培养3d;将子叶的叶面朝下放置于培养基上的愈伤组织诱导率高于叶面朝上的培养方式;子叶诱导胚性愈伤组织的最适培养基配方为MS+6-BA2.0mg/L+KT1.0mg/L+GA31.0mg/L;诱导需要在黑暗条件下启动,进行7d暗培养,而生长分化于光照16h/d和黑暗8h/d条件下培养7d;继代3次得到的胚性愈伤组织最多;最适生根培养基配方为1/2MS+IBA0.3mg/L。体细胞胚胎诱导率最高为25%。胚性愈伤组织经进一步培养发育为成熟的体细胞胚胎,生根成为完整植株。     

Meristem culture and micropropagation of a variety of ginger (Zingiber officinale Rosc.) with a high yield of oleoresin

Summary

Meristems of ginger with or without leaf primordia were induced to form shoots on three-quarter strength Murashige-Skoog’s (MS) medium containing sucrose 6%, coconut milk (CM) 20%, ascorbic acid (AA) 100 mg l^?1, glutamine (GL) 400 mg l^?1, activated charcoal (AC) 250 mg l^?1, 6-benzylaminopurine (BAP) 0.5 mg l^?1, indolebutyric acid (IBA) 0.4 mg l^?1 and agar 0.8%. Meristem-derived shoots exhibited consistent multiplication on three-quarter strength MS medium containing sucrose (3%), AA (100 mg l^?1), AC (100 mg l^?1), BAP (4–5 mg l^?1) and agar (0.8%). Liquid media (agitated or static) were less effective than a solid (agar-gelled) medium for micropropagation. Kinetin and naphthalene acetic acid (NAA) incorporated at various levels (0.01–0.8 mg l^?1) with or without added BAP and IBA neither improved plantlet formation nor enhanced shoot multiplication. The in vitro plants were successfully established in vivo and the rhizome yield was comparable with that of plants grown by conventional methods.     

Rejuvenation influences indirect organogenesis from leaf explants of Pomegranate (Punica granatum L.) ‘Kandhari Kabuli’

Sequential subculturing leads to a gradual physiological change in cells that may be termed ‘rejuvenation’. The effect of repetitive subculturing on callus induction and shoot regeneration from leaf explants of Punica granatum L. ‘Kandhari Kabuli’ were investigated. Surface-sterilised leaves were cultured on 1.0× Murashige and Skoog (MS) medium supplemented with 4.0 mg l^–1 α-naphthaleneacetic acid (NAA) and 2.0 mg l^–1 6-benzyladenine (BA) for callus induction. Shoots were regenerated from callus on 1.0× MS medium supplemented with 1.5 mg l^–1 BA, 0.5 mg l^–1 kinetin, and 0.25 mg l^–1 NAA. Subculturing of callus onto fresh medium maintained the rate of shoot formation and substantially increased the production of shoot buds up to the second subculture. Following further subculture passages, a lower shoot regeneration potential from callus was observed. A maximum shoot bud induction from callus of 63.9% was observed at the second subculture passage. The rate of multiplication of in vitro shoots increased until the fourth subculture, then became constant. Similarly, in vitro rooting of micro-shoots increased up to the third subculture, followed by a decline during further subculturing.     

Factors affecting the efficacy of agar-based substrates for the study of tomato pollen germination

Summary

The present work examines the effects of substrate additions and incubation conditions on in vitro germination of tomato pollen on semi-solid, agar-based substrates with the ultimate aim of defining a standard semi-solid substrate for tomato pollen germination studies. Partial replacement of sucrose by polyethylene glycol (PEG-6000) for osmotic regulation of the substrate significantly increased pollen germination of both the low temperature-susceptible F₁ cultivar ‘Dombito’, and the more temperature-resistant cv.‘Supermarmande’, to a level that varied according to the season of pollen harvest. In contrast, partial substitution of sucrose by mannitol was inhibitory to an extent that depended on the final concentration of mannitol in the medium. The optimum pH for germination was 6.5 and the optimum incubation temperature was 15°C. Among the vitamins (riboflavin, thiamine, pyridoxine, niacin, pantothenic acid, p-aminobenzoic acid), amino acids (glutamic acid, aspartic acid), casein hydrolysate, plant growth regulators (gibberellic acid, indole-3-acetic acid, indole-3-butyric acid, indole-3-propionic acid, 1-naphthaleneacetic acid, kinetin) and flavonoids (naringin, myricetin, naringenin, quercetin, rutin) tested, a significant increase in pollen germination was induced only by the flavonoids quercetin and myricetin and, to a lesser extent, by indole-3-acetic acid. In contrast, both kinetin and naringenin were inhibitory, as were both amino acids at 100 mg l^–1 and casein hydrolysate (at 1.0 – 10.0 g l^–1). On the basis of the above results, we conclude that, for both tomato cultivars tested, the most suitable semi-solid substrate among those examined was that containing 10% (w/v) sucrose, 15.1% (w/v) PEG-6000 and 1.5% (w/v) agar, with the possible addition of 5 mg l^–1 quercetin, or 5 mg l^–1 myricetin.     

Effects of chlorocholine chloride and prohexadione-Ca on rhizome growth and lateral bud production in Iris germanica L.

Iris germanica L. is a popular perennial flower worldwide, but its use is limited in China due to an inadequate availability of propagules. To accelerate rhizome growth and lateral bud production using plant growth retardants (PGRs), chlorocholine chloride (CCC; at 1,500 or 3,000 mg l^?1) or prohexadione-Ca (at 700 or 1,500 mg l^?1) were applied to uniform plants of I. germanica. Time-course measurements of changes in morphogenesis, carbohydrate metabolism, total soluble protein (TSP) concentrations, and endogenous phytohormone concentrations in rhizomes were conducted to test the efficacy of CCC or prohexadione-Ca for increasing rhizome growth and lateral bud production. The results showed that both PGRs increased the fresh weights of rhizomes at 2, 4, and 6 weeks after treatment (WAT). Overall, 700 mg l^?1 prohexadione-Ca was most effective at promoting the formation of lateral buds which increased by 183.5% at 12 WAT relative to untreated control plants. Concentrations of sucrose and starch in PGR-treated rhizomes increased at 2, 4, and 6 WAT, while a decline was observed by 12 WAT. TSP concentrations increased during rhizome enlargement, then decreased during lateral bud germination after prohexadione-Ca treatment. In general, concentrations of endogenous phytohormones, such as gibberellins, indole-3-acetic acid, jasmonic acid, and zeatin riboside, decreased significantly in rhizomes at 4 WAT, then increased at 12 WAT. Our study indicated that prohexadione-Ca promoted rhizome growth and the accumulation of sucrose and starch before summer dormancy, then significantly accelerated the production of lateral buds.     

In vitro vegetative multiplication of Fuchsia hybrida

Rapid development of axillary buds from shoot-tips and nodes of 18 cultivars of Fuchsia hybrida was obtained on solid Murashige and Skoog medium with BAP (1 mg l^?1 and an auxin (0.1 mg l^?1). NAA as the auxin appeared to be more active than IAA or IBA. Vegetative shoots were subsequently isolated and developed up to 15 supplementary axillary shoots on the same solid medium. Agitated and non-agitated liquid media of the same composition were less effective. One-cm long shoots could be rooted in 20 days in the presence of IBA before being transferred to soil.     

Effects of nitrogen and phosphorus nutrition on the growth of asparagus seedlings

N was applied at 50, 100 or 150 mg l^?1 in factorial combination with P at 7.5, 15 or 22.5 mg l^?1 to asparagus seedlings. There were 6 successional harvests. N and P increased shoot dry weight by increasing mean dry weight and number of shoots. Increasing P had no effect on shoot growth at 50 mg l^?1 N. N increased root dry weight (crown and roots) by increasing root number, whereas P decreased root dry weight due to a decrease in mean root dry weight. N increased total plant dry weight, but P had no effect. N and P increased the partitioning of dry weight to the shoots, while partitioning to the roots increased with time. Plant analysis revealed that 2.6–2.7% N and 0.29–0.36% P, on a dry-weight basis, were present in the shoots at the later harvests with the higher concentrations of N and P. 100–150 mg 1^?1 N in combination with 15 mg l^?1 P produced a seedling suitable for transplanting into commercial fields at 6 weeks from emergence.