De novo in vitro shoot morphogenesis from shoot tip-induced callus cultures of Gymnema sylvestre (Retz.) R. Br. ex Sm

BACKGROUND: Gymnema sylvestre is a medicinal woody perennial vine known for its sweetening properties and antidiabetic therapeutic uses in the modern and traditional medicines. Its over-exploitation for the therapeutic uses and to meet the demand of pharmaceutical industry in raw materials supply for the production of anti-diabetic drugs has led to considerable decline in its natural population. RESULTS: An efficient system of shoot bud sprouting from nodal segment explants and indirect plant regeneration from apical meristem-induced callus cultures of G. sylvestre have been developed on Murashige and Skoog (MS) medium amended with concentrations of cytokinins. Of the three growth regulators tested, N6-benzylaminopurine (BAP) was the most efficient and 2.0 mg L-1 gave the best shoot formation efficiency. This was followed by thidiazuron (TDZ) and kinetin (Kin) but, most of the TDZ-induced micro shoots showed stunted growth. Multiple shoot formation was observed on medium amended with BAP or TDZ at higher concentrations. The produced micro shoots were rooted on half strength MS medium amended with auxins and rooted plantlets acclimatized with 87% survival of the regenerates. CONCLUSIONS: The developed regeneration system can be exploited for genetic transformation studies, particularly when aimed at producing its high yielding cell lines for the anti-diabetic phytochemicals. It also offers opportunities for exploring the expression of totipotency in the anti-diabetic perennial vine.

Use of Palm Male Inflorescence and River-Sand as Acclimatization Substrate for Plantain (Musa sp.) Cultivars

The objective of this work was to investigate the use of Palm Male Inflorescence (PMI) and river-sand as substrate for the acclimatization of plantain. Plantlets from three plantain cultivars (Batard, Ebanga and French Clair) were obtained after 16 weeks of tissue cultures and the plantlets were subjected to routine acclimatization under screen house conditions using two different substrates mixed in different ratios (100% Sand, 100% PMI, 75% PMI, 60% PMI and 50% PMI). The experiment was arranged in a completely randomized design with ten (10) replications; each replicate consisting of one micro-pot. The different substrates used significantly influenced the performance of the cultivars. The best medium for acclimatization for French Clair was 60% PMI in terms of percentage survival of plantlets (96.88%), plantlet height (6.03 cm), diameter (0.60 cm), number of leaves (4.42 leaves), leaf area (20.23 cm2), leaf emergence rate (1.64), number of roots (7.70 roots), and root length (18.86 cm). Ebanga plantlets had the best results with 75% PMI in terms of percentage survival of plantlets (96.88%), plantlet height (6.18 cm), diameter (0.62 cm), number of leaves (4.39 leaves), leaf area (20.48 cm2), leaf emergence rate (1.76), and total fresh weight (10.05 g). Meanwhile with Batard cultivar, 50% PMI was the best substrate in terms of percentage survival of plantlets (96.88%), plantlet height (4.41 cm), diameter (0.55 cm), number of leaves (4.55 leaves), leaf area (12.96 cm2), leaf emergence rate (1.55), and number of roots (5.73 roots). This study clearly show that PMI can be a viable substrate to use with sand in plantlet acclimatization; however, the different plant cultivars had optimal result at different proportions of PMI.

Effect of Arbuscular mycorrhizal fungi (AMF) and Azospirillum on growth and nutrition of banana plantlets during acclimatization phase.

The effect of the co-inoculation of arbuscular mycorrhizal fungi (AMF) and Azospirillum on micro-propagated banana seedlings development during their adaptation phase was determined. At the time of transplanting, banana seedlings were inoculated with an indigenous mycorrhizal inoculum containing 10 spores/g at four doses: 0, 50, 100 and 200 g. Seventy days after fungal inoculation, 20 ml of Azospirillum in four concentrations (0, 106, 107 and 108 CFU/ml) were applied. Finally, after 98 days from the start of the experiment a second dose (40 ml) of Azospirillum in the concentrations mentioned above was inoculated. Plants were harvested 5 months after transplanting and the growth and nutritional parameters were evaluated. The analysis of the data showed that banana plants co-inoculated with 200 g of AMF and 1.5E8 CFU/ml of Azospirillum presented greater development, an increase of 7 times in height, 4 times in perimeter, 16 times in leaf area, 12 times in aerial biomass, and 8 times in root biomass relative to control plants. The results achieved were due to synergism between fungus-bacteria when inoculated at higher doses, with lower doses stimulating growth is minimal. The co-inoculation in high doses demonstrates adequate support and cooperative effect between HMA and Azospirillum crops. In addition, co-inoculation promotes optimal nutritional status because microorganisms allowed plants achieve greater absorption of phosphorus and nitrogen relative to those treated with single inoculation and the control.

RAPD analysis of genetic diversity of Dioscorea opposita germplasm resources from Jiangxi and genetic stability of their plantlets / 中草药

Objective: To analyze the genetic diversity of 11 varieties of Dioscorea opposita germplasm resources from Jiangxi, and detect the genetic stability of the tissue cultured plantlets. Methods: RAPD-PCR was applied. Results: Twenty primers were screened out, 238 bands were amplified, 234 of them were polymorphic bands, the average amplified polymorphic bands were 11.7, and the ratio of polymorphic bands was 98.32%. The genetic similarity coefficient was 0.575 6-0.970 6 and the average similarity coefficient was 0.723 1. According to the results of UPGMA clustering analysis, the 11 D. opposita germplasm resources from Jiangxi province were divided into five categories: The first class included D. bulbifera. The second class included D. opposita. The third class included D. opposita Nancheng 1, D. opposita, D. opposita, D. opposite, and D. opposita, which showed that D. opposita 1, D. opposita, D. opposita and D. opposite, had closer relationship with D. opposita came from Wenxian County, Jiaozuo City, Henan province. The fourth class included D. opposita, D. opposita Nancheng 2 and D. opposita. The fifth class included D. opposita. The RAPD analysis results of plantlets subcultured for six times by in vitro rapid propagation method of stem segment with a bud of D. opposita L. germplasm resources from Jiangxi province (treatment group) and their seedlings germinated from microtuber (control group) also showed that 11 yam varieties were spread out 126-179 identifiable bands and each variety appeared 5-18 mutation bands, which indicated plantlets subcultured for six times by in vitro rapid propagation method of stem segment with a bud of D. opposita. germplasm resources from Jiangxi province existed somaclonal variation, but their genetic similarity coefficient compared with their seedlings germinated from microtuber was still relatively high, illustrating that their genetic traits were still relatively stable and did not affect their genotype. Conclusion: The experimental results can provide the reliable basis for introduction breeding, resource improvement, variety identification, germplasm conservation, and their plantlet culture of D. opposita germplasm resources from Jiangxi province.

Studies on Calli Mediated Multiple Shoot Regeneration from Rumex vesicarius L.

Rumex vesicarius L is a valuable potent medicinal herb, which belongs to family Polygonaceae. It possesses antimicrobial, anti- inflammatory, antidiarrhoeal and antioxidant properties. An attempt to study the In vitro callus induction and regeneration of plantlets from calli of leaf and nodal segments as explants has been achieved. Initially mature seeds were excised from plants grown in the departmental garden of KL University. The sterilized seed explants were inoculated aseptically to the solid basal SH media without any growth regulators for seed germination. Effective plantlets observed after 1 week of culture inoculation under maintained controlled conditions. From these In vitro plantlets, leaf and nodal segments were taken as explants for this study. These explants were inoculated on SH medium supplemented with different concentrations of BA (0.5-5.0mg/l) and 2, 4-D (0.5-3.0mg/l) for callus induction and multiple shoot regeneration. 90% of callus induction was observed on media containing BA 4.0mg/l and efficient multiple shoot induction (96.6%) was observed on media containing BA 1.0mg/l from leaf explants. 90% of callus induction and 85% of multiple shoot induction observed on media containing BA 2.0mg/l from nodal explants. Roots were induced from In vitro shoots on SH medium supplemented with 1mg/l IBA after 1 week. Leaf explants were more regenerative with 96.6% response compared to nodal explants 85%. Finally these In vitro regenerated plantlets were hardened, acclimatized and successfully transferred to the field. This protocol will be useful for mass multiplication of plantlets and maintenance of germplasm throughout the year.

In vitro callus induction and plantlet regeneration of Achyranthes aspera L., a high value medicinal plant

Objective:To study callus induction from different explants (internode, leaf, root) and in vitro plantlets propagation from medicinally important plant Achyranthes aspera L. Methods:Sterilized explants were prepared by using 0.1%HgCl2 and 0.5%Bavistin and callus was obtained when cultured onto Murashige Skoog’s (MS) medium by using different concentrations and combination of 2,4-D, NAA, BAP, IAA, IBA with 3%sucrose and 0.8%agar. Induced callus was immediately transferred to MS medium containing at different concentrations of phytohormones for shootlets and rootlets induction respectively. Results:Sterilization treatment of 0.1%HgCl2 for 2-3 min and Bavistin 0.5%for 10-12 min showed the highest percentage of asepsis and survival rate. Maximum induction of callus was obtained from a combination of 2.0 mg/L 2,4-D and 0.5 mg/L NAA from leaf. Highest shootlets number (4.83±0.17) and length (3.8±0.16) cm were observed on full strength MS medium when fortified with BAP 4.0 mg/L and KIN 0.5 mg/L. Concerted efforts of BAP 2.0 mg/L and NAA 0.5 mg/L on full strength MS medium showed highest leaf number (6.77±0.94). In vitro raised shoots were allowed to root on different strengths of MS medium fortified with IAA and IBA at different concentrations. Experimentally, 3.0 mg/L IBA was enabled to induce maximum rootlets number (10.0±9.82) on full strength MS medium. Afterwards, regenerated shoots with well developed roots were successfully subjected to hardening process and were acclimatized. The survived plantlets showed 66.67%survival frequency without any morphological abnormality. Conclusions: The results demonstrated that different explants were good source of callus induction, morphology analysis as well as indirect plantlets regeneration.

In vitro mutation induced by ethylmethane sulfonate in Stevia rebaudiana and SRAP identification of salt-tolerant mutants / 中草药

Objective: The in vitro mutation system of Stevia rebaudiana induced by ethylmethane sulfonate (EMS) was established, and the salt-tolerant mutants were identified by SRAP. Methods: S. rebaudiana plantlets were inoculated on MS media containing NaCl with different concentration to screen the salt-tolerant critical concentration. Plantlets were treated with EMS at different concentration and for different time periods, and EMS mutagenized stems were inoculated on MS medium containing critical NaCl concentration to screen the tolerant variants by SRAP markers. Results: The critical salt concentration of S. rebaudiana plantlets was 1.0%, and the suitable concentration and time of EMS were 0.8%-1.0% and 8-10 h. Among the screened 41 S. rebaudiana tolerant mutants by SRAP molecular markers, four were mutated at DNA level, and the mutation rate was 9.76%. Conclusion: The in vitro mutagenesis system of S. rebaudiana with EMS has been established, which provides a new breeding way for high-yield salt-tolerant S. rebaudiana.

Dual and Axenic culture of Sclerospora graminicola on Pearl Millet and in vitro raised disease resistant host germplasm.

Mother axis of the infected suppressed ear-head of pearl-millet with embryoids was sliced (2-3 mm) and surface sterilized with 0.2 % Ethyl mercuric chloride followed by repeated washings with sterile antioxidant solution (50 mg/L citric acid, 25 mg/L ascorbic acid and 50 mg/L P.V.P). Sterilized slices were cultured on modified MS medium supplemented with 10 mg/L IAA, 0.5 mg/L Kinetin, 2.5 mg/L 2,4-D, 3.0 mg/L casein hydrolysate, 25 mg /L ascorbic acid and 150 mg/L coconut water. Two types of calli were formed after 10-15 days. Slow growing compact milky coloured was embryogenic and the other hyaline fast-growing was non-embryogenic. The fungus grew on the callus after 20-25 days of inoculation. It grew axenically on the surface of the medium after sometime. Slow growing embryogenic callus was subcultured to auxin free ¾ strength MS medium with 25 mg/L ascorbic acid, 100 ml/L coconut water and 0.5 mg/L Kinetin. Regeneration plantlets were transferred for rooting on ½ strength MS medium containing 1.5 mg/L IBA and 25 mg/L ascorbic acid. The regenerated plantlets with strong root system were transferred to 4 earthen pots. The plantlets raised were subjected for disease resistance by growing them on the medium containing different concentrations of the fungus filtrate. The plantlets showing resistance have been accepted as diseases resistant.

Crescimento in vitro e aclimatização de Cattleya loddigesii Lindley (Orchidaceae) com carvão ativado sob dois espectros luminosos / In vitro growth and acclimatization of Cattleya loddigesii Lindley (Orchidaceae) with actived charcoal in two light spectra

O objetivo deste estudo foi avaliar a adição de concentrações de carvão ativado em meio de cultura ½ MS (com metade da concentração dos macronutrientes) sob dois espectros luminosos para a obtenção de plântulas in vitro de Cattleya loddigesii. Plântulas com aproximadamente 90 dias foram subcultivadas em oito tratamentos, nos quais foi testada a adição ao meio de cultura ½ MS com carvão ativado (0; 0,5; 1,0 e 2,0g L-1) e combinados sob espectro de luz branca e luz vermelha. Após 180 dias da germinação, foram mensurados dados biométricos (raiz e parte aérea), massa fresca e teores de pigmentos fotossintéticos. Em plântulas aclimatizadas em casa de vegetação, foram avaliadas a taxa de sobrevivência após 120 dias. As concentrações de clorofila total, clorofila a e carotenoides foram maiores nos tratamentos sob luz branca, enquanto a luz vermelha influenciou significativamente maior clorofila b, plântulas com menos raízes e de menor comprimento e elevada mortalidade ex vitro. A adição de 2,0g L-1 de carvão ativado ao meio de cultura e o uso de luz branca proporcionaram maior eficiência de desenvolvimento tanto para as culturas in vitro quanto para a sobrevivência ex vitro das plantas.

The aim of this study was to evaluate the effect of different concentrations of activated charcoal in ½ MS (half concentration of macronutrients) culture medium under two light spectra on the in vitro growth of Cattleya loddigesii seedlings. Plantlets with approximately 90 days were subcultured under eight treatments, consisting of different active charcoal concentrations (0; 0.5; 1.0 and 2.0g L-1) in ½ MS medium combined with white and red light spectra. After 180 days of germination, biometric data, fresh weight, and the level of photosynthetic pigments were evaluated. Plantlets acclimatized in a greenhouse were evaluated for survival after 120 days. Total chlorophyll, chlorophyll a, and carotenoid concentrations were higher in treatments under white light, while red light promoted greater chlorophyll b, plantlets with fewer and shorter roots, and high ex vitro mortality. The addition of 2.0g L-1 of active charcoal to the culture medium and the use of white light provided greater development efficiency both on in vitro culture and ex vitro plant survival.

Growth response of maize plantlets inoculated with Enterobacter spp., as a model for alternative agriculture / Respuesta de plántulas de maíz inoculadas con Enterobacter spp. como un modelo de agricultura alternativa

A maize rhizosphere isolate was phenotypically and genotypically characterized and identifed as Enterobacter spp. bacterium. Germinated seeds were inoculated, the plantlets were sown in vermiculite and in soil and grown under laboratory and feld conditions, respectively. The adherence, colonization and plant growth promotion capability of Enterobacter sp. UAPS03001 was evaluated in "Rojo-Criollo" maize under laboratory conditions. Twenty days after inoculation, the treated plantlets showed larger biomass than non-inoculated ones. In feld grown plants, the kernel biomass was also greater in inoculated than in non-inoculated plants. The inoculation of maize sprouts with plant growth- promoting bacteria before their sowing in the feld would be an alternative practice for achieving successful yield in temporal agriculture.

En este trabajo se aisló una bacteria de la rizósfera de maíz, que fue caracterizada mediante métodos fenotípicos y genotípicos e identifcada como Enterobacter sp. UAPS03001. La bacteria fue inoculada en semillas de maíz "Rojo-Criollo" germinadas en forma axénica. Las semillas germinadas e inoculadas se plantaron en vermiculita y posteriormente las plántulas fueron cultivadas en vermiculita o en suelo, para evaluar el efecto promotor del crecimiento vegetal de dicha bacteria, bajo condiciones de laboratorio y de campo. Bajo condiciones de laboratorio, también se evaluó la capacidad de esta cepa para adherirse a las plantas de maíz y colonizarlas. Veinte días después de la inoculación, las plántulas inoculadas mostraron una biomasa mayor con referencia a las no inoculadas. En campo, la biomasa de la mazorca fue también mayor en las plantas inoculadas respecto de las plantas no inoculadas. La inoculación de germinados de maíz con una bacteria promotora del crecimiento vegetal y su posterior transferencia a campo podría ser una práctica alternativa para llevar a cabo una producción exitosa en agricultura de temporal.

Concentration of nutrient solution in the hydroponic production of potato minitubers / Concentração da solução nutritiva na produção hidropônica de minitubérculos de batata

The effect of the nutrient solution concentration on potato plant growth and minituber yield were determined in a sand closed hydroponic system. Minitubers and micropropagated plantlets of the cv. 'Macaca' were used. Treatments were five nutrient solution concentrations at electrical conductivities (EC) of 1.0 (T1), 2.2 (T2), 3.4 (T3), 4.7 (T4) and 5.8dS m-1 (T5). The split plot randomised experimental design was used with three replications. Plants from minitubers produced higher fresh and mean weight of minitubers, shoot dry mass and leaf area index than the micropropagated ones. However, higher dry mass of minitubers was found with micropropagated plantlets compared to minitubers. The concentration of the nutrient solution did not affect minituber number. Increasing the nutrient solution concentration decreased total and minituber dry mass production of micropropagated plantlets and plant growth and minituber production of minituber-originated plants. Low concentration of nutrient solution at an EC of about 1.0dS m-1 can be used in the hydroponic production of potato minitubers of both micropropagated and minituber-originated plants.

Neste trabalho foi determinado o efeito da concentração da solução nutritiva no crescimento e na produtividade de minitubérculos de batata em um sistema hidropônico fechado empregando areia como substrato. Plântulas micropropagadas e minitubérculos foram plantados em 24 de março de 2004. Os tratamentos foram cinco soluções nutritivas com condutividades elétricas (CE) de 1,0 (T1), 2,2 (T2), 3,4 (T3), 4,7 (T4) e 5,8dS m-1 (T5). O experimento foi conduzido em parcelas subdivididas no delineamento inteiramente casualizado com três repetições. Plantas originadas de minitubérculos produziram mais massa fresca total e média de minitubérculos, massa seca da parte aérea e maior índice de área foliar que plantas micropropagadas. Entretanto, maior massa seca dos minitubérculos foi obtida em plantas micropropagadas. A concentração da solução nutritiva não afetou o número de minitubérculos. O aumento da CE reduziu a massa seca total e dos minitubérculos de plantas micropropagadas e decresceu linearmente o crescimento e a produtividade de minitubérculos de plantas oriundas de minitubérculos. Concentrações baixas de solução nutritiva com valores da ordem de 1,0dS m-1 podem ser empregadas na produção de minitubérculos de batata a partir de plântulas micropropagadas e minitubérculos.

Crescimento e percentual de emergência de plântulas de citrumeleiro 'Swingle' em função dos substratos e das doses de corretivo à base de Lithothamnium, após cem dias da semeadura / Growth and percentage of emergence of citrumeleiro \"Swingle\" seedlings in relation to substrates and doses of Lithothamnium based correctives, after a hundred days from 'Swingle'

O experimento foi conduzido no telado do pomar da Universidade Federal de Lavras (UFLA), durante o período de novembro de 2003 a fevereiro de 2004. Realizado com o objetivo de avaliar o efeito de seis diferentes substratos (convencional, Plantmax®, composto I, composto II, vermiculita e húmus), interagindo com três doses do corretivo à base de Lithothamnium, 0 por cento, 5 por cento e 10 por cento (v/v), sobre o crescimento do citrumeleiro 'Swingle', até o momento da repicagem. Foram utilizados tubetes plásticos cônicos com volume de 75 mL, acondicionados em bandejas com capacidade para 198 tubetes. O delineamento experimental utilizado foi blocos casualizados, em esquema fatorial de 6 x 3, sendo os fatores compostos pelos seis diferentes substratos e três diferentes doses do corretivo à base de Lithothamnium, compondo assim, dezoito tratamentos com quatro repetições, sendo cada parcela composta por vinte e duas plântulas. Decorridos cem dias após a semeadura, as plântulas foram retiradas dos tubetes, seccionadas na base do coleto,e registrados os comprimentos da parte aérea e da raiz. Em seguida a biomassa seca da raiz e da parte aérea foram avaliadas. Verificou-se que o húmus e o composto I, isoladamente apresentaram os melhores resultados de crescimento. O corretivo à base de Lithothamnium mostrou ser uma alternativa de incremento nutricional para o crescimento de mudas de citrumeleiro 'Swingle', em condições de pH baixas, e com pouca disponibilidade de Ca e Mg.

This experiment was established aiming at evaluating the effect of six different substrates (conventional, Plantmax®, compost I, compost II, vermiculite, and muck), interacting with three different Lithothamnium doses, 0 percent, 5 percent and 10 percent (v/v), over the growth of citrumelo "Swingle" plantlets, until the moment of chiming. The experiment was conducted within the orchard of the Federal University of Lavras, in the state of Minas Gerais, Brazil, during the period of November 2003 until February 2004. The experimental design used was random blocks, in a factorial squeme of 6x3, with the factors constituted by six different substrates and three different Lithothamnium doses, composing thus, eighteen treatments with four repetitions. Each parcel was composed by twenty two plants. One hundred days after the planting, all the plantlets were taken off the plastic tubes, washed under tap water, and were sectioned in the base of the stem. Thus, the data of the length of each of the parts were recorded. Afterwards, the parts of the plantlets, were placed inside paper bags and transferred to forced air drying hoods, at a temperature of 75° C, during a period of 72 h. The dry weight of the canopy and root system were registered. It was observed that muck and compost I, separately, presented the best results in terms of growth and amount of dry matter. Lithothamnium evidenced as an alternative to increase nutrient availability to the citrumelo growth plantlets, but only when the pH conditions and the availability of Ca and Mg were low.

In vitro regeneration of Didymopanax morototoni

The present study aimed at establishing a complete plant regeneration protocol for Didymopanax morototoni (matchwood), a native Brazilian forest species. Four types of explants (root, shoot, node, and cotyledonary leaves) were obtained from in vitro germinated seeds. In the first step, woody plant medium (WPM) with casein hydrolysate (250 mgL-1 ) and 2,4-D (1.0 and 5.0 mgL-1) were used combined with kinetin (0.1 and 1.0 mgL-1). Twenty days after inoculation, the material was evaluated. Embryogenic calli were split, transferred to expression medium with several combinations of NAA and KIN, and moved to fresh medium after 60 days. Light did not interfere in embryo expression. Somatic embryos were formed either from individual cells or cell clusters. Plantlets were obtained in WPM medium and 10 gL-1 of sucrose with no plant regulator, or using 0.1 mgL-1 BAP and 0.5 mgL-1GA. Plantlets from somatic embryos of D. morototoni developed in 33% of the cases.

O presente estudo visou o estabelecimento de um completo protocolo de regeneração para Didymopanax morototoni (morototó, caixeta) uma espécie florestal nativa do Brasil. Quatro tipos de explantes (raiz, caule, nódulo foliar e folha cotiledonar) foram obtidos a partir de sementes germinadas. Na primeira etapa, meio WPM com caseína hidrolisada (250 mgL-1) e 2,4D (1,0 e 5,0 mgL-1) foram usados em combinação com cinetina (0,1 ou 1,0 mg L-1). Vinte dias depois de inoculado, o material foi avaliado. Calos embriogênicos foram divididos e transferidos para meio de expressão com várias combinações de ácido naftaleno-acético e cinetina, e repicados a cada 60 dias para meio novo. A luz não interferiu na expressão embriogênica. Embriões somáticos foram formados ou de células individuais ou de agregados de células. As plântulas foram obtidas no meio WPM com 10 g L-1 de sacarose e sem reguladores de crescimento ou com 0,1 mg L-1 de Benzil-adenina e 0,5 mg L-1 de giberelina. O desenvolvimento das plântulas a partir de embriões somáticos de D. morototoni foi alcançada em 33% dos casos.

Rapid propagation technology of Dioscorea bulbifera virus-free plantlets / 中草药

Objective Effects of different factors on proliferation and rooting of stems with a bud were studied by using Dioscorea bulbifera as test material to optimize the rapid propagation system of D.bulbifera virus-free plantlets.Methods Plant tissue culture method was used in shoot tip culture and rapid propagation study,and RT-PCR method was used in virus detection of virus-free plantlets.Results The best proliferation medium of D.bulbifera stems with a bud was MS+KT 2 mg/L+6-BA 1 mg/L+NAA 0.5 mg/L;The best sucrose and agar concentration of D.bulbifera stems with a bud was 30 g/L and 0 g/L,respectively;The best rooting medium of D.bulbifera stems with a bud was 1/2 MS+IBA 0.1 mg/L+NAA 0.5 mg/L+PP_(333) 1 mg/L;The best transplanting matrix of regeneration plantlets from D. bulbifera stems with a bud was perlite-vermiculite(2:1);The best PP_(333) concentration of D.bulbifera regeneration plantlets for transplanting was 50 mg/L.Conclusion The rapid propagation system of D. bulbifera virus-free plantlets is established successfully for the first time,which provides a technological basis for factory production of D.bulbifera virus-free plantlets.

Botanical characters of test-tube plantlets and seed plantlets in Bupleurum chinense / 中草药

Objective To provide a scientific basis for the creation and promotion of the rapid propagation of excellent Bupleurum chinense and study on botanical characters of test-tube plantlets and seed plantlets in B.chinense.Methods B.chinense with high effective medicinal ingredients and great stability of genetic characters was selected for rapid propagation,and some of botaninal characters between test-tube plantlets and seed plantlets were compared.Results The tiller number,the branch stem number,and the inflorescence number of the test-tube plantlets were significantly greater than those in seed plantlets.Conclusion Through the rapid propagation and the large-scale cultivation of test-tube plantlets of B.chinense,much more roots and seeds are obtained comparing with those of seed plantlets,while maintaining the excellent traits of B.chinense.