Assessment of factors affecting micropropagation and ex vitro acclimatization of Nyctanthes arbor-tristis L

Rapid differentiation of multiple shoots was observed in 94% of nodal explants of one year old Nyctanthes arbor-tristis L. plants. Shoot bud induction and multiplication took place on Murashige and Skoog (MS) medium supplemented with two cytokinins, i.e. Benzyladenine (BA) or Kinetin (Kn) either alone or in combination with different auxins, indole-3-butyric acid (IBA), indole-3-acetic acid (IAA) or α-naphthalene acetic acid (NAA). Between different media, pH levels and growth regulators tried, the optimum condition for maximum regenerative response was obtained on MS + Kn (2.5 μM) + N AA (0.5 μM) media at 5.8 pH, forming cultures with 23.26 ± 0.89 number of shoots and 6.36 ± 0.80 cm shoot length after 8 weeks of culture. Histological sections confirmed the formation of multiple buds from nodal explants. Rooting was achieved ex vitro by dipping the basal ends of microshoots in 200 μM IBA for 30 min followed by their transplantation in sterile soilrite. The plantlets with well-developed shoot and root system were successfully established in garden soil and grown outside in a greenhouse with a 80% survival rate.     

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)...     

Effect of Cytokinins and Cytokinin Antagonists on in vitro Cultured Gypsophila paniculata L.

This study deals with the effects of two cytokinins [kinetin (Kin) and N-(2-chloro-4-pyridyl)-N-phenylurea (CPPU)] and cytokinin antagonists [2-chloro-4-cyclobutyl-amino-6-ethylamino-1,3,5-triazine (ACK1) and N-(4-pyridyl)-O-(4-chlorophenyl)carbamate (ACK2)] in concentration of 1 μM on in vitro cultured Gypsophila. The application of Kin and CPPU stimulated bud opening and increased fresh and dry masses. Cytokinin antagonists reduced the number of sprouted buds and bud fresh and dry masses. In plants treated with CPPU the chloroplasts possessed well developed membrane system, which covered almost the entire chloroplasts volume. In ACK2 treated plants, the plastid apparatus in each cell was represented by two types of chloroplast in which the inner membrane system was differently organized. Cell wall adjacent chloroplasts possessed structure similar to the controls. In inner located chloroplasts part of thylakoids were semi-concentrically arranged and partially destructed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Physical factors increased quantity and quality of micropropagated shoots of Cannabis sativa L. in a repeated harvest system with ex vitro rooting

In Vitro Cellular & Developmental Biology - Plant - Micropropagation is a preferred method to propagate clean, clonal stock plants. Subculture is labor intensive and costly. In vitro hedging...     

UDP-Glucuronosyltransferase activity is correlated to saponin production in Gypsophila paniculata root in vitro cultures

UDP-Glucuronosyltransferase (UGT, EC 2.4.1.17) activity was detected in excised root cultures of Gypsophila paniculata. The UGT activity (up to 0.8 nkat mg^–1 protein) correlated to the total saponin content (2 mg g^–1 dw) during the exponential phase of the batch culture. This gives rise to the hypothesis of a direct relationship in the biosynthetic pathway regulation between the enzyme activity and the saponin biosynthesis.     

生物反应器在满天星快繁中的应用

利用2.5L柱状气升式生物反应器采用接触法进行了满天星(Gypsophila paniculata)增殖培养。结果表明:外植体接种密度(每个反应器接种外植体个数)为35比密度为25和45更有利于满天星的增殖生长,平均每个生物反应器内可获得252株生长健壮的不定苗;光强为90μmol.m-2.s-1对满天星增殖培养最有利,过高的光强对不定芽和株高有抑制作用;通过小孔隙(15μm)的多孔喷头注入0.1vvm空气有利于满天星增殖培养。     

重瓣满天星植株再生及其复壮和移栽

重瓣满天星 (Gypsophila paniculata)花蕾在附加有不同激素的 MS培养基上 (0～ 3mg/L6 - BA,0～ 1mg/L NAA,0～ 1mg/L 2 ,4 - D)均可长出再生植株 ,其中以 MS+6 - BA1mg/L效果最佳。试管苗复壮以 OM +6 - BA 1mg/L +2 ,4 - D 1mg/L效果最好。经复壮的试管苗较粗壮 ,但直接移植仍不能成活 ,必须在加盖旋松的培养瓶中进行溶液培养。2周后 ,小苗根系进一步发育扩展 ,然后逐步揭盖 ,锻炼幼苗抗干能力 ,移栽成活率可达 80 %。     

生物反应器在满天星快繁中的应用

利用2.5 L柱状气升式生物反应器采用接触法进行了满天星(Gypsophila paniculata)增殖培养。结果表明: 外植体接种密度(每个反应器接种外植体个数)为35比密度为25和45更有利于满天星的增殖生长, 平均每个生物反应器内可获得252株生长健壮的不定苗; 光强为90 mmol.m-2.s-1对满天星增殖培养最有利, 过高的光强对不定芽和株高有抑制作用; 通过小孔隙(15 mm)的多孔喷头注入0.1 vvm空气有利于满天星增殖培养。     

Improved in vitro rooting in liquid culture using a two piece scaffold system

Plant tissue culture techniques have been used to propagate horticultural crops at a commercial scale for more than three decades. However, due to the high cost it is generally only used for high value crops. To increase production efficiency and make micropropagation viable for a wider range of species, new approaches to address key steps of the process with high labor inputs need to be evaluated. For this study, a two‐piece scaffold system was designed, prototyped using 3D printing, and tested to physically hold plants upright thereby facilitating liquid based rooting. This system was evaluated with Malus domestica, Betula lenta, and Musa sp. using static liquid culture as well as rocker based temporary immersion system and compared to rooting in semi‐solid based medium as is commonly practiced. Significantly, earlier rooting was observed in all three species in liquid when compared to semi‐solid culture system, and plants cultured in liquid on the rocker generally performed better than those in static liquid. In addition to quicker, more uniform rooting, reducing labor requirements, and preventing root damage. This newly designed system is simple, easy to use, will help to improve efficiency, and reduce the cost of micropropagation.     

满天星试管苗与其玻璃化苗的RAPD指纹图谱分析

采用分离群体分组分析法(BSA),用100个随机引物对满天星的正常苗和玻璃化苗进行RAPD分析的结果表明,7个随机引物扩增出多态性差异条带。再用上述7个引物分别对试管苗及其玻璃化苗个体进行DNA的PCR扩增的结果显示,引物J20在2种苗中出现差异条带。     

Rapid in vitro multiplication and ex vitro rooting of Malus zumi (Matsumura) Rehd

Micropropagation system of Malus zumi was optimized by studying the influence of plant growth regulators and culture conditions. The axillary buds were used for mutiplication of in vitro shoot culture on agar Murashige and Skoog (1962) (MS) medium with combination of 1 mg l⁻¹ BAP, 0.5 mg l⁻¹ NAA or 0.5 mg l⁻¹ IAA or 0.5 mg l⁻¹ IBA under 16 h photoperiod. The shoot growth in culture was not significantly affected within a broad range (5.0–7.0) of initial medium pH. The highest shoot (13) was obtained on medium containing 1.0 mg l⁻¹ BAP and 0.5 mg l⁻¹ IAA. Well-developed shoots, 35–50 mm in length, were successfully rooted ex vitro at 86.3% by a 2-h-treatment with aqueous solution containing MS salts and 100 mg l⁻¹ IBA prior to their planting in growing substrate composed of soil and vermiculite (1:1 v/v). The survival rate of transplantation reached 88.0% when transferred to field condition.     

Development of a novel in vitro rooting culture system for the micropropagation of highbush blueberry (Vaccinium corymbosum) seedlings

Plant Cell, Tissue and Organ Culture (PCTOC) - Here, we established a novel, efficient, and cost-effective rooting culture system for highbush blueberry (Vaccinium corymbosum). This method...     

Uptake,accumulation and phytoextraction efficiency of cesium in Gypsophila paniculata

Abstract

To evaluate the phytoextraction efficiency of Gypsophila paniculata from Cs-contaminated soils and analyze the mechanism of Cs accumulation in G. paniculata, we analyzed the characteristics of Cs bioaccumulation and subcellular distribution, in addition to its chemical forms in the plant under hydroponic conditions. The results showed that total Cs content in the aboveground parts and the entire plant were as high as 6137.32?mg·kg^?1?dry weight and 7338.49?mg·kg^?1?dry weight, respectively, after 17?days in the 50?mg·L^?1 Cs treatment. The BCF was between 2.35 and 3.38. The TF was between 1.00 and 2.46 in G. paniculata. Subcellular distribution of Cs in the plant was as follows: soluble fraction?>?cell wall?>?organelles. Inorganic Cs (F-ethanol) and water-soluble Cs (F-dH2O) were the main types of Cs in G. paniculata. Further studies show that the phytoextraction efficiency can reach 10.30–11.91% planting a season of G. paniculata under potted conditions. The results suggested that G. paniculata, a perennial, drought-tolerant herb, was a high-accumulator of Cs, which may have potential uses in phytoremediation of Cs-contaminated soil.     

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.     

Factors affecting induction and development of in vitro rooting in apple rootstocks

Shoots of apple rootstocks raised in vitro were transferred to various rooting media to study the effect of different factors on root initiation and development. Various concentrations of indole-3-butyric acid (IBA) initiated rooting but maximum rooting percentage was found with 2.0 and 2.5 mg l(-1) of IBA in M7 and with 1.0 mg l(-1) of IBA in MM106. The drawback was that the roots were thick, short and with profuse callus. The presence of activated charcoal (AC) in the rooting medium improved the rooting quality but reduced the rooting percentage in both the rootstocks. In high auxin dip of 70, 80 and 90 mg l(-1) IBA for 2, 2 and 1 hr showed 75-85 per cent rooting in M7, but lacked reproducibility of the results. Whereas in MM106, 66 - 70 % rooting was achieved with 70 mg l(-1) of IBA dip for 3 h. Root induction in shoots in IBA containing liquid medium (LM) in dark for few days and root elongation in IBA--free medium in light proved most effective. On the other hand, continuous light treatment showed reduced rooting. Reduction of MS salts and sucrose in root elongation medium showed decreased rooting. Plantlets from two--stage rooting procedure showed more rapid growth and satisfactory survival during hardening of plants and on transfer to field.     

In vitro micropropagation and ex vitro acclimation of Bupleurum kaoi-an endangered medicinal plant native to Taiwan

Summary This study reports an improved protocol for in vitro-shoot multiplication and ex vitro acclimation of Bupleurum kaoi, an endangered medicinal herb. Nodal segments were cultured in half-strength Murashige and Skoog (MS) basal medium supplemented with different concentrations of benzyladenine (BA) and kinetin. The presence of 0.25 mg l⁻¹ BA induced the highest number of shoots per explant after 8 wk of culture. Although BA was more effective than kinetin on shool multiplication, it induced hyperhydric shoots at all concentrations tested. The use of dispense paper (DP) instead of aluminum foil (AF) for container closure was found to reduce hyperhydricity and improve ex vitro acclimation. The best survival rate (61%) was obtained when plantlets were grown in MS basal medium containing 0.5 mg l⁻¹ indole-3-butyric acid and 0.1–0.2 mg l⁻¹ α-naphthaleneacetic acid using DP as container closure. Leaves of the plant treated with AF6 (two layers of AF as container closure and 6 wk of incubation) lacked epicuticular wax and possessed larger stomata, higher stomata density, and fewer functional stomata compared to those of plants treated with AF2+DP4 (two layers of AF for 2 wk, then replaced AF by three layers of DP for 4wk) and ex vitro-acclimated plantlets.     

Micropropagation and ex vitro rooting of pistachio (Pistacia vera L.)

Plant Cell, Tissue and Organ Culture (PCTOC) - An effective pistachio (Pistacia vera L.) micropropagation system was developed involving rapid axillary bud proliferation and ex vitro rooting. The...     

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)     

In vitro culture of Gypsophila paniculata L. and random amplified polymorphic DNA analysis of the propagated plants

A protocol is established for regeneration of the economically important cut flower plant, Gypsophila paniculata L., using shoot tips explants. Multiple shoots were obtained on Murashige and Skoog medium fortified with 0.5 mg dm⁻³ each of α-naphthaleneacetic acid and 6-benzyladenine. Addition of 10 g dm⁻³ agar promoted shoot proliferation and reduced the degree of shoot vitrification. Transfer to 3 mg dm⁻³ indole⁻³-butyric acid containing medium produced optimum root initiation and development. The produced plants as well as intact plants were subjected to the random amplified polymorphic DNA (RAPD) analysis. Using 9 primers, the total number of amplification products generated by polymerase chain reaction was 142 bands (15.7 bands per primer), of which 7.74 % showed polymorphism. The analysis of bands recorded, showed 92.25 % similarity. The results indicated that very low variation at the DNA level occurred during in vitro culture of Gypsophila.     

Improved in vitro rooting of Prunus dulcis Mill. cultivars

A highly reproducible system was developed for efficient rooting of cultivars Boa Casta (BC) and Peneda and a BC seedling-derived clone (BC VII) of almond (Prunus dulcis Mill.). Twenty-four accessions derived from the clone BC VII and subjected to various in vitro culture treatments were screened. The long induction pre-treatment (LIP, 5 d), the brief induction pre-treatment (BIP, 16 h) and the hormonal shock by short dipping in hormone solution (1 min), were tested. BIP was the only that allowed rooting of cultivars. In BC VII, it induced high rooting frequencies (47–100 %) when using a solution of 0.4 mM indole-3-butyric acid solidified with 2 g dm⁻³ gellam gum for 16-h. The response to the auxin type was variable depending on the cultivar and the root induction pre-treatment used. Root number was significantly different between the two cultivars and BC VII. Root length was significantly higher when using 0.005 mM IBA in LIP but this concentration induced apical necrosis. The improved acclimatization procedure for up to 4 weeks increased the survival to 45 %. The initiation and development of adventitious roots were proved to be asynchronous.