Morphogenic Response of Leaf and Petiole Explants of Broccoli Using Thidiazuron

Broccoli (Brassica oleracea var. italica) is an important nutritionally rich vegetable cole crop grown in the world. Environmental stress, pests, and diseases cause enormous yield losses because of a limited gene pool. Genetic manipulation is becoming an important method for broccoli improvement. The objective of present study was to evaluate the potency of thidiazuron (TDZ) as a plant growth regulator in evoking morphogenic responses in leaf and petiole explants of broccoli. An efficient, reproducible, and high frequency plant regeneration protocol has been standardized in broccoli cv. Solan green head. Leaf and petiole explants were cultured on Murashige-Skoog (MS) medium, supplemented with a wide range of TDZ concentrations. The following treatments were designed for efficient in vitro shoot regeneration: TDZ alone, TDZ with adenine, TDZ with naphthalene acetic acid (NAA), and TDZ with indole acetic acid (IAA). Among the 36 combinations of growth regulators used, the highest percentage of leaf explants producing shoot (89.25%) was recorded on MS medium containing 1.0 μM TDZ and 0.107 μM NAA. The multiple shoot regeneration response of petiole explant producing shoots (91.55%) was obtained on MS medium containing 2.0 μM TDZ and 0.107 μM NAA. Shoot multiplication and elongation were obtained on the same medium. For root regeneration in in vitro regenerated shoots, different concentrations of NAA were applied. High frequency (100%) root regeneration response with healthy and vigorous roots was observed on MS medium supplemented with 0.54 μM NAA. The regenerated plantlets with well-developed shoots and root system were transferred to pots containing cocopeat and successfully acclimatized. We recommend 1.0 μM TDZ with 0.107 μM NAA and 2.0 μM TDZ and 0.107 μM NAA combinations for adventitious shoot regeneration from leaf and petiole explants in broccoli cv. Solan green head respectively. This is the first report on high frequency organogenesis from leaf and petiole explants of broccoli cv. Solan green head using thidiazuron.     

Mass multiplication of Bixa orellana L. through tissue culture for commercial propagation

To reduce the time period for in vitro regeneration in annatto (Bixa orellana L.), a highly efficient two-stage plant regeneration protocol had been developed that can be used commercially. Different types of explants: nodal shoot tips, shoot tips and single nodes from in vitro grown seedlings were inoculated onto the Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of plant growth regulators. Highest number of shoot buds was obtained when nodal shoot tip explants were inoculated onto MS medium supplemented with 31.1 μM N⁶-benzyladenine (BA) and 14.7 μM phenylacetic acid (PAA). PAA in combination with BA exhibited a synergistic effect on shoot multiplication and elongation. Sub-culturing of the shoots onto the MS medium supplemented with BA (13.3 μM) and PAA (7.3 μM) produced elongated shoots. Elongated shoots when inoculated onto the MS medium supplemented with 4.9 μM indole-3-butyric acid (IBA) produced optimal rooting. The rooted plantlets were hardened and their field survival rate after 6 weeks time was 73%.     

TDZ预处理及激素诱导对小麦茎尖丛生芽形成的影响

为了给遗传转化提供更好的受体,从而降低离体芽转化直接成苗出现的嵌合体现象,以4个普通小麦品种(系)为材料,研究了小麦籽粒细胞分裂素TDZ(噻二唑苯基脲)浸泡预处理及激素对小麦茎尖丛生芽形成的影响。结果表明,TDZ浸泡预处理降低了小麦胚萌发率,但显著提高了小麦茎尖丛生芽的分化,在TDZ最佳浓度5mg·L-1时,4种供试小麦材料的丛生芽诱导率和平均芽个数均达到最高,其中扬麦18最优,丛生芽诱导率和平均芽个数分别达到了50.9%和2.82个;萌发培养基中的TDZ浓度为4mg·L-1时,小麦茎尖丛生芽的诱导率和平均芽个数最高;诱导培养基中TDZ和6-BA(6-苄基脲嘌呤)浓度均为3mg·L-1时,诱导小麦丛生芽的效果最佳,并且TDZ诱导效果优于6-BA;细胞分裂素与生长素配合诱导时,在TDZ/IAA(吲哚乙酸)为6、6-BA/2,4-D(2,4-二氯苯氧乙酸)为60时,诱导效果较好,但低于细胞分裂素单独诱导的效果,且随着比值的进一步增大,其诱导效率急速下降;同时基因型对小麦茎尖丛生芽诱导有重要影响,供试的4个材料中,扬麦18的丛生芽诱导率最高,扬麦12次之,西农183、小偃22较差。     

In vitro regeneration from petiole explants of non-toxic Jatropha curcas

Jatropha curcas, a multipurpose shrub has acquired significant economic potential as biodiesel plant. The seeds or pressed cake is toxic due to the presence of toxic substances and is not useful as food/fodder despite having the best protein composition. A simple, efficient, and reproducible method for plant regeneration through direct organogenesis from petiole explants of non-toxic J. curcas was developed using Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ). The best induction of shoot buds (57.61%), and number of shoot buds (4.98) per explant were obtained when in vitro petiole explants were placed horizontally on MS medium supplemented with 2.27 μM TDZ. The Induced shoot buds were transferred to MS medium containing 10 μM kinetin (Kn), 4.5 μM 6-benzyl aminopurine (BA), and 5.5 μM α-naphthaleneacetic acid (NAA) for shoot proliferation and subsequent elongation was achieved on MS medium supplemented with 2.25 μM BA and 8.5 μM IAA. The elongated shoots could be rooted on half-strength MS medium with 15 μM IBA, 11.4 μM IAA and 5.5 μM NAA with more than 90% survival rate.     

Regeneration in Jatropha curcas: Factors affecting the efficiency of in vitro regeneration

Factors influencing in vitro regeneration through direct shoot bud induction from hypocotyl explants of Jatropha curcas were studied in the present investigation. Regeneration in J. curcas was found to be genotype dependent and out of four toxic and one non-toxic genotype studied, non-toxic was least responsive. The best results irrespective of genotype were obtained on the medium containing 0.5 mg L⁻¹ TDZ (Thidiazuron) and in vitro hypocotyl explants were observed to have higher regeneration efficiency as compared to ex vitro explant in both toxic and non-toxic genotypes. Adventitious shoot buds could be induced from the distal end of explants in all the genotypes. The number of shoot buds formed and not the number of explants responding to TDZ treatment were significantly affected by the position of the explant on the seedling axis. Explants from younger seedlings (≤15 days) were still juvenile and formed callus easily, whereas the regeneration response declined with increase in age of seedlings after 30 days. Transient reduction of Ca²⁺ concentrations to 0.22 g L⁻¹ in the germination medium increased the number of responding explants.Induced shoot buds, upon transfer to MS medium containing 2 mg L⁻¹ Kn (Kinetin) and 1 mg L⁻¹ BAP (6-benzylamino purine) elongated. These elongated shoots were further proliferated on MS medium supplemented with 1.5 mg L⁻¹ IAA (indole-3-acetic acid) and 0.5 mg L⁻¹ BAP and 3.01-3.91 cm elongation was achieved after 6 weeks. No genotype specific variance in shoot elongation was observed among the toxic genotypes except the CSMCRI-JC2, which showed reduced response. And for proliferation among the toxic genotypes, CSMCRI-JC4 showed highest number of shoots formed. Among the rest, no significant differences were observed. The elongated shoot could be rooted by pulse treatment on half-strength MS medium supplemented with 2% sucrose, 3 mg L⁻¹ IBA (indole-3-butyric acid), 1 mg L⁻¹ IAA, 1 mg L⁻¹ NAA (α-naphthalene acetic acid) and subsequent transfer on 0.25 mg L⁻¹ activated charcoal medium. The rooted plants could be established in soil with more than 90% success. No significant differences were observed in rooting of shoots in the different toxic genotypes. However, rooting response was reduced in non-toxic genotype as compared to toxic genotypes.     

In vitro plant regeneration from decapitated embryonic axes of Clitoria ternatea L.—An important medicinal plant

In vitro clonal propagation of Clitoria ternatea has been achieved by employing decapitated embryonic axes (DEAs) explants. The explants induced multiple shoots on cytokinin-containing medium. Several cytokinins [6-benzylaminopurine (BAP), 6-furfuryl aminopurine (KIN) and thidiazuron (TDZ)] were assayed. The best response was achieved with 2 mg l⁻¹ BAP in which 100% of cultures produced 6.0 ± 0.14 shoots per explant. MS + 1 mg l⁻¹ gibberellic acid (GA₃) was the most suitable for shoot elongation. Regenerated shoots were rooted in half-strength Murashige and Skoog (MS) medium with 0.2 mg l⁻¹ indole-3-butyric acid (IBA). Plantlets were successfully acclimatized and established in soil, and they were morphologically indistinguishable from the source plant. The plantlets attained maturity and flowered normally. The efficient regeneration protocol reported here provides an important method of micropropagation of this plant. Furthermore, this protocol may be used for genetic transformation of this valuable medicinal plant for its further improvement.     

An improved and efficient micropropagation of Eclipta alba through transverse thin cell layer culture and assessment of clonal fidelity using RAPD analysis

An improved and efficient in vitro regeneration system has been developed for Eclipta alba, a medicinally important plant, through transverse thin cell layer culture (tTCL). The transverse section of the nodal segment of field grown plants was used as tTCL explants for plant regeneration. Shoot multiplication from tTCL nodal explants was influenced by BAP and their interaction with Kin or NAA. MS medium containing 13.2 μM BAP and 4.6 μM Kin was most effective for shoot multiplication from tTCL nodal explants. Upon this medium, percent response for shoot proliferation was 100% with an average of 32.6 shoot buds per tTCL nodal explant. Regenerated shoots from tTCL nodal explants were rooted on the growth regulator free MS medium. The rooted plantlets were successfully acclimatized and established in soil with a survival frequency of 90-100%. Random amplified polymorphic DNA (RAPD) markers were used to evaluate the genetic fidelity of the micropropagated plants. RAPD profile analysis indicated that micropropagated plants were genetically similar to mother plant.     

花生幼叶芽诱导和植株再生研究

从萌发9－10d的花生幼嫩叶片上切取中段作外植体，接种MS＋BA 3mg/L NAA 0.8mg/L AgNO3 2mg/L诱芽培养基，12－14d后产生丛生芽点，芽诱导率达78．9％，平均每外植体产生9个丛生芽。4周后转至MS BA 3mg/L AgNO3 2mg/L诱导芽伸长，诱导生根后获得再生植株。     

TDZ和暗培养对蝴蝶兰叶片不定芽发生的影响

以蝴蝶兰幼嫩花梗诱导的无菌苗叶片为外植体,以MS为基本培养基,研究不同暗培养时间和TDZ浓度对蝴蝶兰叶片诱导不定芽的影响。结果表明：暗培养和TDZ对接种的蝴蝶兰叶片的成活率均具有显著影响,暗培养60 d时,叶片的存活率在90%以上,而在同一暗培养时间条件下,叶片的存活率随着TDZ浓度的增加而上升;在TDZ浓度为3.0 mg/L,暗培养60 d时,蝴蝶兰存活叶片不定芽的诱导率最高,达到93.45%;诱导的不定芽数最多,平均每个外植体诱导的不定芽数为13.22个。综合考虑外植体的成活率、不定芽诱导率和诱导的不定芽数,蝴蝶兰叶片诱导不定芽的最佳条件为：MS＋TDZ 3.0 mg/L,暗培养60 d。     

Somatic embryogenesis and regeneration of five multipurpose cassava landraces extensively integrated in African cropping system

Successful development and adoption of transgenic cassava (Manihot esculenta Crantz) varieties in Africa depend on in vitro regeneration of landraces because of their agronomic value and amenability to genetic modification. The study investigated somatic embryogenesis from axillary bud and immature leaf-lobe explant and regeneration via shoot organogenesis in five cassava landraces. Landraces exhibited significant (P < 0.05) differences in frequencies of primary somatic embryo production, number of embryos per explant, and frequency of secondary embryos. The frequency of primary somatic embryogenesis ranged from 7.8 to 14.5%, whereas the number of embryos per explant varied from 5.3 to 10.6. However, only frequency of primary somatic embryos and number of embryos per explant showed significant (P < 0.05) differences when immature leaf lobe was used as explant. The frequency of primary somatic embryogenesis ranged from 42.7 to 49.2%, whereas number of embryos per immature leaf-lobe explant varied from 9.5 to 15.2 per explant. Secondary embryogenesis was cultivar-independent in the case of immature leaf-lobe explant. Cyclic and green (mature) embryogenesis showed no significant (P < 0.05) landrace differences in both axillary bud and immature leaf-lobe explants. Shoot regeneration from cotyledon of somatic embryos had significant (P < 0.05) landrace differences. The mean shoot-bud formation frequency of axillary bud and immature leaf-lobe explants was 51.4% and 37.2%, respectively. Root formation was efficient, with greater than 70% of shoots forming root in all landraces. Similarly, landrace differences were detected for the survival of acclimatized regenerated plants, with a mean of 94.7% for both explants. In conclusion, somatic embryos were produced from immature leaf and axillary bud explants of the landraces and the embryos were converted to plantlets via shoot organogenesis.     

‘MaxClover’ grazing experiment: I. Annual yields,botanical composition and growth rates of six dryland pastures over nine years

Six dryland pastures were established at Lincoln University, Canterbury, New Zealand, in February 2002. Production and persistence of cocksfoot pastures established with subterranean, balansa, white or Caucasian clovers, and a perennial ryegrass‐white clover control and a lucerne monoculture were monitored for nine years. Total annual dry‐matter (10.0–18·5 t DM ha^?1) and sown legume yields from the lucerne monoculture exceeded those from the grass‐based pastures in all but one year. The lowest lucerne yield (10 t ha^?1 yr^?1) occurred in Year 4, when spring snow caused ungrazed lucerne to lodge and senesce. Cocksfoot with subterranean clover was the most productive grass‐based pasture. Yields were 8·7–13·0 t DM ha^?1 annually. Subterranean clover yields were 2·4–3·7 t ha^?1 in six of the nine years which represented 26–32% of total annual production. In all cocksfoot‐based pastures, the contribution of sown pasture components decreased at a rate equivalent to 3·3 ± 0·05% per year (R² = 0·83) and sown components accounted for 65% of total yield in Year 9. In contrast, sown components represented only 13% of total yield in the ryegrass‐white clover pastures in Year 9, and their contribution declined at 10·1 ± 0·9% per year (R² = 0·94). By Year 9, 79% of the 6.6 t ha^?1 produced from the ryegrass‐white clover pasture was from unsown species and 7% was dead material. For maximum production and persistence, dryland farmers on 450–780 mm yr^?1 rainfall should grow lucerne or cocksfoot‐subterranean clover pastures in preference to ryegrass and white clover. Inclusion of white clover as a secondary legume component to sub clover would offer opportunities to respond to unpredictable summer rainfall after sub clover has set seed.     

Mycorrhizal fungi and growth and development of micropropagated Scutellaria integrifolia plants

Scutellaria species have been used in many traditional medical systems and is well known among the Native American tribes as a strong emmenagogue and as a female medicinal herb. The inoculation of arbuscular mycorrhiza fungi (AMF) into the roots of micropropagated plantlets could help not only mass propagate these species, but also help grow in marginal, phosphorus deficient soils. Leaves, shoot apices, and nodal segments from wild as well as from greenhouse-grown plants were used to initiate cultures in Murashige and Skoog (MS) medium supplemented with cytokinins benzyladenine (BA), kinetin, thidiazuron (TDZ), naphthalene acetic acid (NAA), and indole butyric acid (IBA) Among all the explants tested for shoot bud induction, only shoot tips and nodal explants were responsive. Explants swelled and became rough on the surface at the end of 3-week incubation with many green shoot buds. Two to 3 weeks after transfering to rooting media with or without IBA, all shoots developed roots. In vitro raised plants were acclimatized in a mist chamber and transferred to 6-l containers in the greenhouse to study the role of AMF on plant growth and development. Inoculation with AMF, showed positive effects on plant growth, particularly root development compared with the control plants. Among the five AMF strains tested, S3004 increased plant height and fresh weights of shoot, root, and seed.     

High-efficiency regnerationin vitro from potato petioles with intact leaflets

The shoot/plantlet regenerationin vitro of seven potato (Solanum tuberosum L.) cultivars from petioles with intact leaflets was assessed using six treatment combinations-a basal medium with or without silver thiosul-phate (STS) or thidiazuron (TDZ) at two concentrations (2 or 0.5 mg/l) of the indoleacetic acid (IAA). The basal medium consisted of Murashige and Skoog (MS) salts and vitamins supplemented with 3 mg/1 6-benzylaminopurine, and 1 mg/1 gibberellic acid, 30 g/l sucrose, and 7.0 g/l PHYTOAGAR. Two full sets repeats and one partial set repeat of independent experiments were conducted and all produced similar results. Silver thiosulphate decreased the regeneration frequency and number of shoots per callus. No significant changes were observed with thidiazuron. Regeneration rates of (100% ) with up to 20 shoots/plantlets per callus were achieved at 2 mg/1 IAA with cultivars Désirée, Kennebec, Niska, and Lenape. These cultivars still showed high regeneration rates (87%–98% ) on media with 0.5 mg/1 IAA, and good regeneration rates were also achieved by the other three cultivars (48%, 94%, and 50% for Chieftain, Russet Burbank, and Shepody, respectively). Even with the single medium protocol (0.5% IAA without thiosulphate or thidiazuron), Désirée, Lenape, and Niska exhibited a regeneration rate of 98%. The use of petiole-with-leaflet explants could be ideal for the regeneration step of potato genetic transformation protocol because of their high regeneration efficiency and their small cut surface area forAgrobacterium elimination after co-incubation.     

In vitro shoot regeneration and chromosome doubling in 2X and 3X potato clones

The possibility of scaling ploidy levels up and down offers several advantages for genetic studies and breeding work in potato. Shoot regeneration and chromosome doubling in plants regenerated fromin vitro cultures were investigated in 4 diploid (2n=2x=24) and 2 triploid (2n=3x=36) clones. Internode and leaf expiants were taken from plants propagated eitherin vitro as shoot cultures, orin vivo in a greenhouse. Two regeneration procedures were compared. Regeneration frequencies were significantly higher using a two-step regeneration protocol and from leaf explants, while doubling was more efficient starting fromin vivo grown plants. No doubling was observed in triploid clones. Considering altogether the percentage of regeneration and doubling, and the number of shoots regenerated per explant, the most efficient conditions were considered leaf explants taken fromin vivo grown plants, and cultured according to a two-step regeneration protocol.     

Differential growth response of Rytidosperma species (wallaby grass) to phosphorus application and its implications for grassland management

Rytidosperma species (formerly Austrodanthonia) are native grasses common in temperate grasslands of southern Australia. Nine Rytidosperma species, Lolium perenne and Bromus hordeaceus were grown as microswards in pots in a glasshouse, and their growth response to six levels of applied P was measured. Shoot yield differed up to twofold between the highest‐ and lowest‐yielding Rytidosperma species. Some Rytidosperma species were slow growing with minimal ability to respond to increased soil P availability. However, three species, Rytidosperma duttonianum, Rytidosperma racemosum and Rytidosperma richardsonii, had a similar shoot yield to L. perenne. Species that grew well at high P also grew well at low P, except B. hordeaceus, which was the lowest‐yielding species at low P, but had among the highest yields at high P. No species showed evidence of P toxicity. The species exhibited a range in critical external P requirement (i.e. amount of P applied for 90% maximum yield). Among the fast‐growing Rytidosperma species, R. richardsonii was notable because it had a low critical external P (16·3 mg P pot^?1) and high agronomic P‐use efficiency (94·1 g DW g^?1 P applied). In contrast, R. duttonianum had a higher critical external P requirement (22·4 mg P pot^?1) and lower agronomic P‐use efficiency (85 g DW g^?1 P applied). It was concluded that it is important to know which Rytidosperma species are present in a grassland to understand how it may respond to P fertilization. The results help to explain the diverse opinions expressed about the productivity of pastures containing Rytidosperma species.     

Highly efficient in vitro regeneration of the industrial oilseed crop Crambe abyssinica

A highly efficient regeneration protocol for oilseed crop Crambe abyssinica has been developed using hypocotyls as explants in this study. Crambe is a potential engineering oilseed crop for industrial purposes as it contains 55-60% erucic acid in its oil and, more importantly, it does not outcross with any food oil seed crops. However, the low regeneration frequency with the currently available protocols is still a limiting factor for genetic modification of Crambe. In this study, we investigated the effects of N-source, C-source, AgNO₃, cultural conditions as well as the concentration and combination of plant growth regulators (PGR) on the regeneration frequency of C. abyssinica. The results showed that all these factors, especially the N-source and PGR concentrations and combinations, played an important role in shoot regeneration. Among all the factors tested, the combination of using hypocotyls from C. abyssinica cv. galactica, the Lepiovre basal medium supplemented with 16 g l⁻¹ glucose, 0.5 g l⁻¹ AgNO₃, 2.2 mg l⁻¹ thidiazuron (TDZ), 0.5 mg l⁻¹ α-naphthaleneacetic acid (NAA), 2.5 g l⁻¹ Gelrite, seeds germinated in dark for 3 days and explants cultured in light, gave the best regeneration frequency (over 95%). The results also suggest that reducing the content of NH₄⁺ or keeping a suitable NO₃⁻/NH₄⁺ ratio in the regeneration medium would be crucial to Crambe shoot regeneration.     

In vitro reproduction in the annual pasture legumes subterranean clover (Trifolium subterraneum L.) and French serradella (Ornithopus sativus Brot.)

Pasture legumes are important components of both mixed farming rotations and permanent pastures in temperate climates. Breeding of two widely sown pasture legumes, subterranean clover (Trifolium subterraneum L.) and French serradella (Ornithopus sativus Brot.), is constrained by the long generation cycle, typically enabling only one generation per year. We hypothesized manipulation of culture medium and conditions would enable the development of a laboratory‐based protocol for in vitro reproduction in subterranean clover and French serradella. In vitro flowering and viable seed set was induced from both species. For subterranean clover, the most effective treatment was culturing on modified MS medium with 1 μm kinetin and 0·1 m sucrose under a 100 μmol m^?2 s^?1 light intensity and continuous photoperiod. For French serradella, culture on a hormone‐free B5 medium with 5 mm NH₄Cl and 0·1 m sucrose under a 100 μmol m^?2 s^?1 light intensity and 20 h photoperiod was optimum. It is expected this technique will have application in accelerating generation turnover within breeding programs, for the study of factors influencing flowering in pasture legumes, and for the propagation of valuable yet enfeebled plants such as embryo‐rescued hybrids.     

二穗短柄草成熟胚再生体系建立及农杆菌介导转化研究

为建立二穗短柄草组织培养及遗传转化体系,以二穗短柄草BD21-3成熟胚为外植体,对成熟胚愈伤诱导、分化以及农杆菌侵染条件进行了研究。结果表明,在含有2.5mg·L~(-1) 2,4-D的培养基上,愈伤组织出愈率最高为93.83%;在含有0.2mg·L~(-1) KT的分化培养基上,分化率最高为38.18%;对二穗短柄草胚性愈伤组织农杆菌转化和GUS染色结果表明,侵染的过程中农杆菌菌液浓度OD_(600)为0.6、侵染时间为5min时转化率最高。     

Large scale in vitro propagation of Stevia rebaudiana (bert) for commercial application: Pharmaceutically important and antidiabetic medicinal herb

Stevia rebaudiana is a valuable medicinal plant species and it is being used for the treatment of diabetes. Currently, there is a high demand for raw material of this medicinal herb due to ever increasing diabetes disorder among the population. In order to meet the increased demand an efficient in vitro propagation of S. rebaudiana was established. Nodal explants collected from the field were cultured on MS basal medium fortified with different concentrations of BAP (0.5-3.0 mg/l) and KIN (0.5-3.0 mg/l) individually for shoot bud induction. In vitro derived nodal buds were cultured on MS medium supplemented with different concentrations (0.5-3.0 mg/l) of BAP and KIN for multiple shoot bud regeneration. In the second experiment, in vitro derived buds were placed on MS medium supplemented with different concentrations of BAP (0.5-3.0 mg/l) in combination with 0.5 mg/l IAA or IBA or NAA for shoot bud multiplication. The highest frequency (94.50%) of multiple shoot regeneration with maximum number of shoots (15.69 shoots/explant) was noticed on MS medium supplemented with 1.0 mg/l BAP. For large scale plant production, in vitro derived nodal bud explants were cultured on MS medium fortified with 1.0 mg/l BAP, in which about 123 shoots/explant were obtained after three subcultures on the same media composition. Elongated shoots (>2 cm) dissected out from the in vitro proliferated shoot clumps were cultured on half-strength MS medium containing different concentrations of NAA (0.1-0.5 mg/l) and/or MS medium fortified with various concentrations (0.5-2.0 mg/l) of auxins (NAA, IAA and IBA) for root induction. Highest frequency of rooting (96%) was noticed on half-strength MS medium augmented with 0.4 mg/l NAA. The rooted plantlets were successfully transferred into plastic cups containing sand and soil in the ratio of 1:2 and subsequently established in the greenhouse. The present in vitro propagation protocol would facilitate an alternative method for rapid and large-scale production of this important antidiabetic medicinal plant.     

大豆子叶节高频愈伤组织诱导及其植株再生

以萌发7d的大豆子叶节切断及其切片为外植体,研究了萌发培养基、不同激素及其配比对大豆子叶节愈伤组织诱导与分化的影响.结果表明:当不添加BA时,在子叶节切断处几乎无愈伤组织的产生,只有添加BA时,才能产生愈伤组织.外植体切取方式、愈伤组织诱导培养基中BA和TDZ的浓度配比对愈伤组织诱导率产生很大影响.BA浓度为0.05～...