Bud stem tissue culture of Tulipa edulis / 中草药

Objective: To establish the tissue culture system of callus induction and cluster shoot proliferation with bud stems of Tulipa edulis. Methods: Bud stems were isolated from cooled T. edulis bulbs as explants. The calli were inducted on MS media with different concentration of 6-BA and NAA, and the cultural conditions of shoot differentiation and multiplication were optimized. Results: The optimal medium for callus induction was MS + 6-BA 0.5 mg/L + NAA 2.0 mg/L with a callus inducation rate of 78.54%. After subcultured in original medium, the callus was turned into differentiation medium. The optimal medium for callus differentiation was MS + 6-BA 2.0 mg/L + NAA 0.2 mg/L with a shoot differentiation rate of 66.21%. The optimal medium for the shoot multiplication was MS + 6-BA 0.5 mg/L + NAA 0.2 mg/L, and the proliferation coefficient was 2.48. Conclusion: The media for callus induction, adventitious bud differentiation, and cluster shoot proliferation are optimized. The optimal medium for the culture of T. edulis bud stems is preliminarily established.

Comparison of haploid and doubled haploid sugar beet clones in their ability to micropropagate and regenerate

Background: Haploid plant material is considered as recalcitrant to organogenesis, propagation, and maintenance in vitro. However, sugar beet (Beta vulgaris L.) breeders utilizing doubled haploid (DH) technology in their breeding programs indicate that sugar beet haploids may be cultured in vitro as well as diploids. Thus in this paper the in vitro performance of haploid and the doubled haploid sugar beet of various origin was evaluated. The DHs were derived from haploids by diploidization and twelve such haploid and corresponding DH clone pairs were obtained thus the comparison included haploid and DH clones that had identical allelic composition and differed only in their ploidy level. Results: The genotypes differed in shoot morphology and susceptibility to blackening during culture in vitro, but no significant differences were observed between the haploids and DHs. The micropropagation rate was, on average, higher for the haploids than DHs. Viability of the midrib and petiole explants after a 6-week culture was highly genotype dependent, but not affected by explant ploidy level. However, regeneration efficiency depended on both the genotype and ploidy level. The explants of several haploids regenerated more frequently and developed more adventitious shoots than the corresponding DHs thus overall efficiency was higher for haploids. Conclusions: The results obtained indicate that most of the haploids used in the comparison performed similar to or even better than DHs. This suggests that sugar beet haploid material can be successfully used not only for the production of DHs, but also maintained in vitro and utilized in projects requiring haploid tissues as the source material.

Accumulation of flavonoids and coumarins during adventitious bud growth in Stellera chamaejasme / 中草药

Objective: To investigate the accumulation of flavonoids and coumarins during the growth of adventitious buds in Stellera chamaejasme. Methods: The optimal culture media for the induction and the growth of adventitious buds were screened through tissue culturing. The dynamic accumulation of the total flavonoids and three components of coumarin (umbelliferone, daphnetin, and scopoletin) was analyzed by visible spectrophotometry and high-performance liquid chromatography methods, respectively. Results: The optimal condition for the induction of adventitious bud proliferation was MS + 0.5 mg/L BA + 0.5 mg/L NAA; After 4-week culture, the number of adventitious buds in every tip-bud was 35; The optimal condition for the growth of adventitious bud was MS + 0.3 mg/L BA + 0.1 mg/L NAA; After 4-week culture, the average length of buds was 2.3 cm; The amount of total flavonoids increased to the highest of 0.218 g after 5-week culture of the adventitious buds; The amount of the three components of coumarin increased with culturing time increasing during the growth of adventitious buds, and increase to the highest at week 5. But the contents of the components were lower than those in the wild roots, and the descending order had changed. Conclusion: The accumulation of flavonoids and coumarins in adventitious buds of S. chamaejasme could reach to the highest at week 5 of culture.

Tissue culture of Cecropia glaziovii Sneth (urticaceae): vegetative micropropagation and plant regeneration from callus / Cultura de tecidos de Cecropia glaziovii Sneth (Urticaceae): micropropagação vegetativa e regeneração de plantas via calos

Cecropia glaziovii is a tree with used in Brazilian popular medicine. Methods allowing the clonal propagation of this species are of great interest for superior genotype multiplication and perpetuation. For this reason, we examined the effect of different culture media and different types of explants on adventitious shoot regeneration from callus and buds of C. glaziovii. Leaves, petioles and stipules obtained from aseptically grown seedlings or from pre-sterilized plants were used to initiate cultures. Adventitious shoot regeneration was achieved when apical and axillary buds were inoculated on gelled Murashige & Skoog (MS) medium supplemented with 6-benzylaminopurine alone (BAP) (1.0, 5.0 or 10.0 mg L-1) or combined with -naphthalene acetic acid (NAA) (1.0 or 2.0 mg L-1), after 40 days of culture. Best callus production was obtained after 30 days of petioles' culture on gelled MS medium with 2,4 dichlorophenoxyacetic acid (2,4-D) (5.0 mg L-1) combined with BAP (1.0 mg L-1). Successful shoot regeneration from callus was achieved when MS medium supplemented with zeatin (ZEA) (0.1 mg L-1) alone or combined with 2,4-D (1.0 or 5.0 mg L-1) was inoculated with friable callus obtained from petioles. All shoots were rooted by inoculation on MS medium supplemented with indole-3-acetic acid (IAA) (1.0 mg L-1). Rooted plants transferred to potting soil were successfully established. All in vitro regenerated plantlets showed to be normal, without morphological variations, being also identical to the source plant. Our study has shown that C. glaziovii can be propagated by tissue culture methods, allowing large scale multiplication of superior plants for pharmacological purposes.

Cecropia glaziovii é uma planta lenhosa, popularmente usada no Brasil como medicinal. Métodos que visem a sua propagação clonal podem ser de grande utilidade na preservação de seus genótipos de elite. Foram examinados efeitos de diferentes reguladores de crescimento e explantes na formação de brotações múltiplas a partir de calos e a partir de brotos apicais. Folhas, pecíolos e estípulas obtidas de plântulas assépticas e da esterilização de sementes ou através da esterilização direta dos explantes foram utilizados para iniciar os cultivos. Brotações múltiplas foram obtidas quando brotos apicais ou axilares foram inoculados em meios compostos de sais de Murashige & Skoog (MS), suplementado com apenas 6-benzilaminopurine (BAP) (1,0, 5,0 ou 10,0 mg L-1) ou combinado com ácido -naftaleno acético (ANA) (1,0 ou 2,0 mg L-1), depois de 40 dias. A produção de calos foi obtida após 30 dias, quando pecíolos foram inoculados em meio MS acrescido de acido 2,4 diclorofenoxiacético (2,4-D) (5,0 mg L-1) combinado com BAP (1,0 mg L-1). A regeneração de brotos a partir de calos foi obtida quando meio MS acrescido de apenas zeatina (ZEA) (0,1 mg L-1) ou combinado com 2,4-D (1,0 ou 5,0 mg L-1) foi inoculado com calos friáveis obtidos de pecíolos. As plântulas foram enraizadas em MS suplementado com ácido 3-indol acético (AIA) (1.0 mg L-1). A aclimatação das plântulas enraizadas foi estabelecida pela transferência para vasos contendo substrato orgânico e vermiculita sob umidade relativa 100 por cento. As plântulas regeneradas "in vitro" apresentaram aparência normal e idênticas à planta-mãe. Nosso estudo concluiu que genótipos de elite de C. glaziovii podem ser propagados em larga escala através de métodos "in vitro", proporcionando uma fonte confiável de matéria prima para estudos farmacológicos.

Plant regeneration from cotyledonary explants of Eucalyptus camaldulensis dehn and histological study of organogenesis in vitro

The present work aimed at regenerating plants of Eucalyptus camaldulensis from the cotyledonary explants and describing the anatomy of the tissues during callogenesis and organogenesis processes, in order to determine the origin of the buds. The cotyledonary leaves of E. camaldulensis were cultured in Murashige and Skoog (MS), WPM and JADS media supplemented with 2.7 µM NAA and 4.44 µM BAP. The best results for bud regeneration were obtained on MS and WPM media (57.5 and 55 percent of calluses formed buds, respectively). Shoot elongation and rooting (80 percent) were obtained on MS/2 medium (with half-strength salt concentration) with 0.2 percent activated charcoal. Acclimatization was performed in the growth chamber for 48 h and then the plants were transferred to a soil:vermiculite mixture and cultured in a greenhouse. Histological studies revealed that the callogenesis initiated in palisade parenchyma cells and that the adventitious buds were formed from the calluses, indicating indirect organogenesis.

Este trabalho teve como objetivo a obtenção de plantas de Eucalyptus camaldulensis a partir de folhas cotiledonares e o estudo da anatomia dos tecidos durante a calogênese e organogênese para determinar a origem das gemas. Folhas cotiledonares foram cultivadas em meios de cultura MS, WPM e JADS suplementados com 2,7 µM de ANA e 4,44 µM de BAP. Os melhores resultados para a regeneração de gemas foram obtidos com os meios MS e WPM. Para o alongamento e enraizamento, o meio de cultura MS/2 contendo 0,2 por cento de carvão ativado apresentou-se eficiente para ambas as etapas. A aclimatização foi realizada mediante a abertura dos frascos na sala de crescimento por 48 horas, seguido da transferência para casa-de-vegetação com nebulização intermitente. Estudos histológicos foram conduzidos e revelaram que a calogênese teve início nas células do parênquima paliçádico e que as gemas adventícias formaram-se a partir dos calos, indicando a organogênese indireta.

Selection of Induction Factors for Adventitious Buds of Sarcandra glabra in Vitro / 广州中医药大学学报

Objective To establish the culture technology system for the in-vitro induction of adventitious buds of Sarcandra glabra(Thumb.) Nakai by investigating the induction factors of in-vitro adventitious buds.Methods Stem segments of Sarcandra glabra were cultured as explants.The effects of culture medium with different concentrations of 6-benzylaminopurine(6-BA) and indole butyric acid(IBA) added,lighting time,light intensity,and temperature on the induction of adventitious buds from the explants were observed.Results The medium containing both 6-BA and IBA was more effective to induce shoot formation of Sarcandra glabra than the medium only containing 6-BA or IBA;high concentration of plant hormones would block the growth of the plantlets.The scattering light or faint direct light was suitable for the shoot formation,while dark or strong light would induce the plantlets withered or even dead.Temperature in the range of 25 ℃～30 ℃ was beneficial for the growth of adventitious buds,and the bud induction rate arrived 100% at the temperature of 25 ℃.Conclusion At the temperature of 25 ℃,the faint sunlight or direct light less than 2000 lux,and the medium with 2.5 mg?L-1 6-BA and 0.1 mg?L-1 IBA added can stimulate significantly the adventitious bud formation of Sarcandra glabra,the induction rate up to 82.98%.