Meta-topolin and liquid medium mediated enhanced micropropagation via ex vitro rooting in Vanilla planifolia Jacks. ex Andrews

Plant Cell, Tissue and Organ Culture (PCTOC) - An advanced micropropagation protocol has been developed for the global spice crop Vanilla planifolia using meta-topolin [mT, 6-(3-hydroxybenzylamino)...     

In vitro propagation of Allium tuberosum Rottl. ex. Spreng. by shoot proliferation

Halved shoot bases of Allium tuberosum Rottl. ex Spreng. proliferated both axillary and adventitious shoots on B5 medium (1968) supplemented with either 6-benzylaminopurine (0.5 mg/l) or 1-naphthalene acetic acid (0.1 mg/l) and 2-isopentenyladenine (0.5 mg11). Ia vitro shoots proliferated further numerous shoots upon subculture to fresh medium, and these shoots rooted spontaneously. Plantlets were transplanted successfully to soil and retained the diploid condition of the parents.     

In vitro propagation of Allium tuberosum Rottl. ex. Spreng. by shoot proliferation

Seeds of trifoliate orange (Poncirus trifoliata (L.) Raf.) are sensitive to desiccation, and could not withstand reduction in moisture level below 20%, whereas the excised embryonic axes could be easily desiccated to moisture levels as low as 14% without much loss in viability. Axes could be successfully cryopreserved in liquid nitrogen (–196°C) for eight months. The viable embryonic axes exhibited good growth on modified Murashige and Skoog medium supplemented with 1-Naphthalene acetic acid (NAA) and 6-Benzylaminopurine (BAP). Growth of cryopreserved axes was promoted in the presence of charcoal in the medium allowing for plant recovery.Abbreviations NAA Napthaleneacetic acid - BAP 6-Benzylamino-purine - MS Murashige and Skoog (1962) - LN Liquid nitrogen     

Phenolic compounds profiles during ex vitro acclimatization of micropropagated Hypericum polyanthemum.

Accumulation of benzopyrans and total phenolic compounds were assessed in acclimatized field grown plants of Hypericum polyanthemum, an endemic species of southern Brazil, harvested at different developmental stages. The HPLC analysis of bioactive compounds 6-isobutyryl-5,7-dimethoxy-2,2-dimethylbenzopyran (HP1), 7-hydroxy-6-isobutyryl-5-methoxy-2,2-dimethyl-benzopyran (HP2) and 5-hydroxy-6-isobutyryl-7-methoxy-2,2-dimethyl-benzopyran (HP3) revealed that the three benzopyrans are accumulated both in the vegetative and reproductive parts with maximum contents observed after 18 weeks (in the former) and 20 weeks (in the later) of plant growth (1.92+/-0.085 g % DW and 2.62+/-0.13 g % DW in the vegetative and reproductive parts, respectively). Highest contents of HP1 (1.56+/-0.12 g % DW) and HP2 (0.19+/-0.01 g % DW) were quantified in the green floral buds of the plants, whereas HP3 reached the highest level (1.02+/-0.08 g % DW) in the overblown flowers. The evaluation of total phenolic compounds showed that the vegetative parts accumulated the highest levels of the metabolites (51.93+/-0.67 mg QE (g DW)(-1)) after 16 weeks of plant growth. Considering the reproductive parts, the open flowers accumulated the greatest levels of the bioactive compounds (75.99+/-0.95 mg QE (g DW)(-1)). The results show that H. polyanthemum can be efficiently propagated and acclimatized to produce benzopyrans and other phenolic compounds.     

In vitro propagation of endangered Lippia filifolia mart. and schauer ex schauer

Summary This work describes an efficient micropropagation protocol of Lippia filifolia. Nodal segments cultivation in MS medium supplemented with 6-benzylaminopurine (4.5 μM)/α-naphthaleneacetic acid (NAA; 54nM) induced multiple shoots (in average 27 shoots per explant). Elongated shoots were rooted with NAA (0.11 μM) and they maintained ploidy level of the in vitro produced explants. The basic chromosome number were 2n=2x=24. Regenerated rooted shoots were successfully acclimatized under shading house conditions. This is the first report involving the establishment of a protocol for shoot multiplication and rooting for endangered L. filifolia, contributing for germplasm preservation of this species.     

Micropropagation of Acacia mearnsii from ex vitro material

Multiple shoots were produced from nodal explants, of 30-day-old in vitro grown seedlings and from pretreated 3- and 9-month-old greenhouse grown Acacia mearnsii plants, respectively. Explants were sterilized using 0.1% and 0.2% HgCl2 for 15 min for 3- and 9-month-old explants, respectively. Nodal explants were induced to form multiple shoots when placed on MS medium supplemented with 2.0 mg l –1 benzyladenine. Rooting of these shoots was achieved on MS medium supplemented with 1.0 mg l –1 indole-3-butyric acid. Plantlets were acclimatized in transparent plastic containers under greenhouse conditions with a 90% success rate.     

Factors affecting in vitro shoot proliferation and ex vitro establishment of mangosteen

Shoot proliferation has been achieved in Garcinia mangostana L. using seed explants. Maximum mean number of shoots per explant (16.8) was obtained from cultures on Murashige and Skoog medium supplemented with 40 mM 6- benzyladenine, and 2.5 mM -naphthaleneacetic acid and kept at 30 °C under an 8 hour photoperiod. Cultures on the same medium but supplemented with 2 g l^-1 activated charcoal produced fewer shoots. However, growth of these shoots was more organized and 75% rooting was obtained. Woody Plant Medium was not a suitable medium for shoot proliferation. Ex vitro establishment was best obtained on planting medium consisting of sand, soil and organic material (3:2:1).Abbreviations BA 6-benzyladenine - IBA indole-3-butyric acid - NAA -naphthaleneacetic acid - MS Murashige & Skoog (1962) basal medium - WPM Woody Plant basal medium (Lloyd & Mc Cown 1980)     

How in vitro light affects growth and survival of ex vitro cassava

Cassava is one of the most important food crops in Africa. Meristem culture is an effective method of eliminating viruses and other systemic diseases spread through the vegetative propagation of stems. However, in semi‐arid conditions, survival of ex vitro plants in the field is often disappointing. When an increasing range of light regimes in vitro was provided, the fresh and dry masses more than doubled their values between 29 and 369 mmol s^?1 m^?2 PPFDs. Increases in numbers of senescent leaves and stem thickness were also recorded with increasing PPFD. However, PPFD above 101 mmol s^?1 m^?2 resulted in 30–70% reduction in plant survival, with the thin plants with the smallest fresh and dry masses being the ones with highest survival rates. High light and temperature levels in the greenhouse were also found to be critical for plant survival. It was also shown that transpirational loss from detached leaves and epicuticular wax deposits were not good indicators for predicting survival of ex vitro cassava plantlets during acclimatisation.     

山橙胚的离体培养和植株再生

1植物名称 山橙（Melodinus suaveolens Champ．ex Benth．），又名马骝藤、马骝橙藤、猴子果。     

In vitro propagation of Bambusa nutans Wall. ex Munro through axillary shoot proliferation

This communication describes for the first time an efficient and reproducible protocol for large-scale multiplication of Bambusa nutans. Nodal segments collected from field-grown clumps and cultured on Murashige and Skoog (MS) medium supplemented with 4.4 μM benzylaminopurine (BA) and 2.32 μM kinetin (Kin) gelled with 0.2% gelrite yielded 80% aseptic cultures with 100% bud-break. The in vitro-formed shoots obtained after bud-break were successfully multiplied in MS liquid medium supplemented with 13.2 μM BA, 2.32 μM Kin, and 0.98 μM indole-3-butyric acid (IBA). Sub-culturing of shoots every 3 weeks on fresh multiplication medium yielded a consistent proliferation rate of 3.5-fold. Shoot clusters containing three to five shoots were successfully rooted with 100% success on half-strength MS liquid medium supplemented with 9.8 μM IBA, 2.85 μM indole-3-acetic acid (IAA), 2.68 μM naphthaleneacetic acid (NAA), and 3% sucrose. Plantlets grown in vitro were acclimatized and subsequently transferred to the field. Inter-simple sequence repeat analysis has confirmed the genetic uniformity of the tissue-cultured plants up to 27 passages.     

In vivo and in vitro anti-inflammatory activity of Cryptostegia grandiflora Roxb. ex R. Br. leaves

Background

Despite Cryptostegia grandiflora Roxb. ex R. Br. (Apocynaceae) leaves are widely used in folk Caribbean Colombian medicine for their anti-inflammatory effects, there are no studies that support this traditional use. Therefore, this work aimed to evaluate the effect of the total extract and primary fractions obtained from Cryptostegia grandiflora leaves, using in vivo and in vitro models of inflammation, and further get new insights on the mechanisms involved in this activity.

Results

Ethanolic extract of Cryptostegia grandiflora leaves, and its corresponding ether and dichloromethane fractions, significantly reduced inflammation and myeloperoxidase activity (MPO) in ear tissue of mice treated with 12-O-tetradecanoyl-phorbol-13-acetate (TPA). Histological analysis revealed a reduction of edema and leukocyte infiltration. Complementarily, we demonstrated that extract and fractions reduced nitric oxide (NO•) and prostaglandin E2 (PGE2) production in LPS-stimulated RAW 264.7 macrophages, as well as scavenging activity on DPPH and ABTS radicals.

Conclusions

Our results demonstrated for the first time the anti-inflammatory activity of Cryptostegia grandiflora leaves, supporting its traditional use. This activity was related to inhibition of MPO activity, and PGE2 and NO• production. These mechanisms and its antioxidant activity could contribute, at least in part, to the anti-inflammatory effect showed by this plant.     

Structural effects on Cattleya xanthina leaves cultivated in vitro and acclimatized ex vitro

In vitro orchid micropropagation is efficient biotechnological strategy for conservation and commercial plantlet production. However, micropropagated plantlets generally need to adapt to survive severe changes in humidity, irradiance, and growing medium that accompany the transfer to ex vitro conditions. Such adaptive cellular changes would give insights into the phenotypic plasticity of the model plant Cattleya xanthina (L.) Van den Berg. Therefore, we aimed to evaluate structural changes in the leaves of C. xanthina cultivated in vitro and acclimatized ex vitro using qualitative and quantitative analyses. During acclimatization, we observed a higher accumulation of dry mass, a greater convexity of the outer surface of epidermal cells, an increased deposition of epicuticular waxes, a greater elongation of mesophyll parenchymatic cells, and finally, the presence of chloroplasts with organized thylakoids and well-developed grana. Stomatal density was not changed. Furthermore, a gradual acclimatization allows this species the best adaptation to a new environment.     

始于不定根诱导的沙氏鹿茸草离体培养和植株再生

1植物名称沙氏鹿茸草（Monochasma savatieri Franch.ex Maxim.）,又名绵毛鹿茸草。2材料类别（1）生长在浙江省磐安县新渥镇东侧山坡上的野生绿色植株,2006年10月带土采掘后植于塑料杯中运回贵阳待用（由贵州宏奇药业有限公司提供）。（2）同年6月采自上述地点的种子（提供者同上）。     

Effect of polyamines on in vitro anther culture of Citrus clementina Hort. ex Tan

The improvement of the induction rate in Citrus anther culture is important for taking practical advantage of the haploid potential in breeding. The influence of polyamines on anther culture of Citrus clementina, cv Nules, with particular attention to the free, soluble and insoluble-conjugated polyamine levels, has been investigated. Putrescine, spermidine and putrescine plus spermidine, were added to the standard induction medium. Before culture, spermidine was the most abundant among the free polyamines detected in anthers. The exogenous supply of either putrescine or spermidine, either independently or combined, effected greater uptake and accumulation of polyamines. The addition of 2 mM spermidine to the medium stimulated gametic embryogenesis in clementine Nules, whereas putrescine did not influence embryo production. Regenerants were mostly tri-haploids; a few doubled-haploids and no haploid plants were obtained.     

美丽豹子花的离体快繁和瓶内结球

1 植物名称 美丽豹子花（Nomocharis basilissa Farrer．ex W．E．Evans）。 2材料类别 鳞片切片。 3培养条件 （1）鳞片切片诱导培养基：MS＋BA0．1mg·L^-1（单位下同）＋IBA1．0＋糖60g·L^-1；（2）无菌鳞片增殖培养基：MS＋KT0．5＋NAA0．1＋糖30g·L^-1；     

In vitro and ex vitro Selection of Potato Plantlets for Resistance to Early Blight

Reaction to the culture filtrate of Alternaria solani (Sorauer) was used as an indicator in an in vitro screening test to select lines with decreased field susceptibility to early blight from a population of 1000 putative mutants. Plantlets of cv. ‘Desirée derived from irradiated callus of potato were inoculated in vitro with a culture filtrate of A. solani (Sorauer). Of the 45 lines selected and subsequently evaluated under conditions of natural infection in the greenhouse six showed lesser degrees of early blight infection than the cv. Desirée control. The six lines selected in the greenhouse retained lower degrees of infection during 2 years of field trials.     

安娜竹芋的离体培养与快速繁殖

1 植物名称安娜竹芋(Calathea veitchiana J.G.Veitch ex J.D.Hooker). 2 材料类别新生侧芽. 3 培养条件启动培养基:(1)MS+6-BA 6.0 mg·L-1(单位下同)+NAA 0.05+0.6%卡拉胶+3%白砂糖;增殖继代培养基:(2)MS+6-BA 3.5+KT 2.0+0.6%卡拉胶+3%白砂糖:生根培养基:(3)1/2MS+NAA0.5+0.5%卡拉胶+2%白砂糖.     

In vitro and ex vitro rooting of Siratia grosvenorii, a traditional medicinal plant

Since a decade, the large-scale commercial production of Siratia grosvenorii plantlets is being practiced through in vitro culture of its microcuttings, but it has some drawbacks such as handling of plantlets, low transplant-survival rate, development of massive callus, low yield after transplantation, etc. An experiment has been conducted to improve the prevailing technique as well as to develop a new ex vitro technique to overcome these drawbacks. Several concentrations of naphthalene acetic acid (NAA) (0–4.0 mg/l) have been tried with the MS (Murashige and Skoog in Physiol Plant 15:473–479, 1962) basal medium containing 3% (w/v) sucrose and 4.0 g/l agar, out of which 0.1 mg/l NAA was found best in terms of smaller diameter of callus and maximum rooting and transplant survival rate. Further, use of perlite instead of agar medium also showed possibilities for future research on commercial-scale plantlet production. Ex vitro rooting technique was found superior to the in vitro one as plantlets developed through this method had lateral roots without any callus at the base of microcuttings, just like the natural root system and of course with higher root length, rooting rates, and transplant survival rate compared to the in vitro developed plantlets. Further, this technique is economical in terms of labor and time saving and gives rise to vigorous plants which ultimately bring higher yields and profits.     

Allelopathic effects of Cyperus rotundus extract in vitro and ex vitro on banana

Allelopathic effects of Cyperus rotundus on banana ‘Grande Naine’ were studied in vitro as well as ex vitro. To study the allelopathic effects in vitro, the Cyperus extract was added to the multiplication medium during preparation before adjusting the pH. Of the four concentrations, 0.2 and 0.6% decreased the shoot multiplication and shoot length but 1 and 2% extract completely inhibited the shoot multiplication but induced rooting in 50 and 28% shoots, respectively. For the ex vitro studies, the Cyperus extract was added to the hydroponic medium during the hardening of in vitro raised banana plantlets. The extract of 1 and 2% concentrations decreased the shoot and root length, number of leaves and new roots, fresh weight, total chlorophyll and protein. The fluorescence intensity ratio was successively increased resulting in decreased photosynthesis.     

Characteristics of strawberry plants propagated by in vitro bioreactor culture and ex vitro propagation method

Reproducible protocol for regeneration of complete plantlets from ‘Bounty’ strawberry (Fragaria ananassa Duch.), using a combination of gelled medium and bioreactor system, has been standardized. Sepals, leaf discs, and petiole halves produced multiple buds and shoots when cultured on semi solid‐gelled medium containing 4 μM thidiazuron (TDZ) for 4 wk followed by transferring in liquid medium containing 2 μM TDZ in a bioreactor system and cultured for another 4 wk. TDZ induced shoot proliferation at 0.1 μM in the bioreactor system but inhibited shoot elongation. TDZ‐induced shoots were elongated and rooted in vitro on gelled medium containing 2 μM zeatin. Such bioreactor‐derived tissue culture (BC) plantlets obtained from sepal explants were grown ex vitro and compared with those propagated by tissue culture on gelled medium (GC) and by conventional runner cuttings (RC), for growth, morphology, anthocyanin content, and antioxidant activity after three growth seasons. The BC and GC plants produced more crowns, runners, leaves, and berries than the RC plants although berry weight per plant did not differ significantly. BC and GC plants produced berries with more anthocyanin contents and antioxidant activities than those produced by the RC plants. However, intersimple sequence repeat (ISSR) marker assay produced a homogenous amplification profile in the tissue culture and donor control plants confirming the clonal fidelity of micropropagated plants. In vitro culture on TDZ and zeatin‐containing nutrient media apparently induced the juvenile branching characteristics that favored enhanced vegetative growth with more crown, runners, leaf, and berry production.